Gonadotropin-releasing hormone receptor antagonists and methods relating thereto

ABSTRACT

GnRH receptor antagonists are disclosed which have utility in the treatment of a variety of sex-hormone related conditions in both men and women. The compounds of this invention have the structure:                    
     wherein A, Q, R 1 , R 2 , R 3a , R 3b , R 4 , R 5 , R 6  and n are as defined herein, including stereoisomers, prodrugs and pharmaceutically acceptable salts thereof. Also disclosed are compositions containing a compound of this invention in combination with a pharmaceutically acceptable carrier, as well as methods relating to the use thereof for antagonizing gonadotropin-releasing hormone in a subject in need thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Patent Application No. 60/239,683, filed Oct. 11, 2000, and U.S. Provisional Patent Application No. 60/177,933, filed Jan. 25, 2000, both of which applications are hereby incorporated by reference in their entirety.

STATEMENT OF GOVERNMENT INTEREST

Partial funding of the work described herein was provided by the U.S. Government under Grant No. R43-HD38625 provided by the National Institutes of Health. The U.S. Government may have certain rights in this invention.

TECHNICAL FIELD

This invention relates generally to gonadotropin-releasing hormone (GnRH) receptor antagonists, and to methods of treating disorders by administration of such antagonists to a warm-blooded animal in need thereof.

BACKGROUND OF THE INVENTION

Gonadotropin-releasing hormone (GnRH), also known as luteinizing hormone- releasing hormone (LHRH), is a decapeptide (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH₂) that plays an important role in human reproduction. GnRH is released from the hypothalamus and acts on the pituitary gland to stimulate the biosynthesis and release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). LH released from the pituitary gland is responsible for the regulation of gonadal steroid production in both males and females, while FSH regulates spermatogenesis in males and follicular development in females.

Due to its biological importance, synthetic antagonists and agonists to GnRH have been the focus of considerable attention, particularly in the context of prostate cancer, breast cancer, endometriosis, uterine leiomyoma, and precocious puberty. For example, peptidic GnRH agonists, such as leuprorelin (pGlu-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), have been used to treat such conditions. Such agonists appear to function by binding to the GnRH receptor in the pituitary gonadotropins, thereby inducing the synthesis and release of gonadotropins. Chronic administration of GnRH agonists depletes gonadotropins and subsequently down-regulates the receptor, resulting in suppression of steroidal hormones after some period of time (e.g., on the order of 2-3 weeks following initiation of chronic administration).

In contrast, GnRH antagonists are believed to suppress gonadotropins from the onset, and thus have received the most attention over the past two decades. To date, some of the primary obstacles to the clinical use of such antagonists have been their relatively low bioavailability and adverse side effects caused by histamine release. However, several peptidic antagonists with low histamine release properties have been reported, although they still must be delivered via sustained delivery routes (such as subcutaneous injection or intranasal spray) due to limited bioavailability.

In view of the limitations associated with peptidic GnRH antagonists, a number of nonpeptidic compounds have been proposed. For example, Cho et al. (J. Med. Chem. 41:4190-4195, 1998) discloses thieno[2,3-b]pyridin-4-ones for use as GnRH receptor antagonists; U.S. Pat. Nos. 5,780,437 and 5,849,764 teach substituted indoles as GnRH receptor antagonists (as do published PCTs WO 97/21704, 98/55479, 98/55470, 98/55116, 98/55119, 97/21707, 97/21703 and 97/21435); published PCT WO 96/38438 discloses tricyclic diazepines as GnRH receptor antagonists; published PCTs WO97/14682, 97/14697 and 99/09033 disclose quinoline and thienopyridine derivatives as GnRH antagonists; published PCTs WO 97/44037, 97/44041, 97/44321 and 97/44339 teach substituted quinolin-2-ones as GnRH receptor antagonists; and published PCT WO 99/33831 discloses certain phenyl-substituted fused nitrogen-containing bicyclic compounds as GnRH receptor antagonists.

While significant strides have been made in this field, there remains a need in the art for effective small molecule GnRH receptor antagonists. There is also a need for pharmaceutical compositions containing such GnRH receptor antagonists, as well as methods relating to the use thereof to treat, for example, sex-hormone related conditions. The present invention fulfills these needs, and provides other related advantages.

SUMMARY OF THE INVENTION

In brief, this invention is generally directed to gonadotropin-releasing hormone (GnRH) receptor antagonists, as well as to methods for their preparation and use, and to pharmaceutical compositions containing the same. More specifically, the GnRH receptor antagonists of this invention are compounds having the following general structure (I):

including stereoisomers, prodrugs and pharmaceutically acceptable salts thereof, wherein A, Q, R₁, R₂, R_(3a), R_(3b), R₄, R₅, R₆, and n are as defined below.

The GnRH receptor antagonists of this invention have utility over a wide range of therapeutic applications, and may be used to treat a variety of sex-hormone related conditions in both men and women, as well as a mammal in general (also referred to herein as a “subject”). For example, such conditions include endometriosis, uterine fibroids, polycystic ovarian disease, hirsutism, precocious puberty, gonadal steroid-dependent neoplasia such as cancers of the prostate, breast and ovary, gonadotrophe pituitary adenomas, sleep apnea, irritable bowel syndrome, premenstrual syndrome, benign prostatic hypertrophy, contraception and infertility (e.g., assisted reproductive therapy such as in vitro fertilization). The compounds of this invention are also useful as an adjunct to treatment of growth hormone deficiency and short stature, and for the treatment of systemic lupus erythematosis. The compounds are also useful in combination with androgens, estrogens, progesterones, and antiestrogens and antiprogestogens for the treatment of endometriosis, fibroids, and in contraception, as well as in combination with an angiotensin-converting enzyme inhibitor, an angiotensin II-receptor antagonist, or a renin inhibitor for the treatment of uterine fibroids. In addition, the compounds may be used in combination with bisphosphonates and other agents for the treatment and/or prevention of disturbances of calcium, phosphate and bone metabolism, and in combination with estrogens, progesterones and/or androgens for the prevention or treatment of bone loss or hypogonadal symptoms such as hot flashes during therapy with a GnRH antagonist.

The methods of this invention include administering an effective amount of a GnRH receptor antagonist, preferably in the form of a pharmaceutical composition, to a mammal in need thereof. Thus, in still a further embodiment, pharmaceutical compositions are disclosed containing one or more GnRH receptor antagonists of this invention in combination with a pharmaceutically acceptable carrier and/or diluent.

These and other aspects of the invention will be apparent upon reference to the following detailed description. To this end, various references are set forth herein which describe in more detail certain background information, procedures, compounds and/or compositions, and are each hereby incorporated by reference in their entirety.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, the present invention is directed generally to compounds useful as gonadotropin-releasing hormone (GnRH) receptor antagonists. The compounds of this invention have the following structure (I):

including stereoisomers, prodrugs and pharmaceutically acceptable salts thereof,

wherein:

Q is a direct bond or —(CR_(8a)R_(8b))_(r)—Z—(CR_(10a)R_(10b))_(s)—;

A is O, S, or NR₇;

r and s are the same or different and independently 0, 1, 2, 3, 4, 5 or 6;

n is 2, 3 or 4;

Z is a direct bond or —O—, —S—, —NR₉—, —SO—, —SO₂—, —OSO₂—, —SO₂O—, —SO₂NR₉—, —NR₉SO₂—, —CO—, —COO—, —OCO—, —CONR₉—, —NR₉CO—, —NR₉CONR_(9a), —OCONR₉— or —NR₉COO—;

R₁ and R₂ are the same or different and independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl, substituted heterocyclealkyl, —C(R_(1a))(═NR_(1b)) or —C(N R_(1a)R_(1c))(═NR_(1b));

or R₁ and R₂ taken together with the nitrogen atom to which they are attached form a heterocycle ring or a substituted heterocycle ring;

R_(3a) and R_(3b) are the same or different and, at each occurrence, independently hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl, substituted heterocyclealkyl, —COOR₁₄ or —CONR₁₄R₁₅;

or R_(3a) and R_(3b) taken together with the carbon atom to which they are attached form a homocyclic ring, substituted homocyclic ring, heterocyclic ring or substituted heterocyclic ring;

or R_(3a) and R_(3b) taken together form ═NR_(3c);

or R_(3a) and the carbon to which it is attached taken together with R₁ and the nitrogen to which it is attached form a heterocyclic ring or substituted heterocyclic ring;

R₄ is higher alkyl, substituted alkyl, aryl, substituted aryl, heterocycle, substituted heterocycle, —COR₁₁, —COOR₁₁, —CONR₁₂R₁₃, —OR₁₁, —OCOR₁₁, —OSO₂R₁₁, —SR₁₁, —SO₂R₁₁, —NR₁₂R₁₃, —NR₁₁COR₁₂, —NR₁₁CONR₁₂R₁₃, —NR₁₁SO₂R₁₂ or —NR₁₁SO₂NR₁₂R₁₃;

R₅ is hydrogen, halogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, alkoxy, alkylthio, alkylamino, cyano or nitro;

R₆ is higher alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl;

R₇ is hydrogen, —SO₂R₁₁, cyano, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl; and

R_(1a), R_(1b), R_(1c), R_(3c), R_(8a), R_(8b), R₉, R_(9a), R_(10a), R_(10b), R₁₁, R₁₂, R₁₃, R₁₄ and R₁₅ are the same or different and, at each occurrence, independently hydrogen, acyl, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl or substituted heterocyclealkyl;

or R_(1a) and R_(1b), R_(8a) and R_(8b), R_(10a) and R_(10b), R₁₂ and R₁₃, or R₁₄ and R₁₅ taken together with the atom or atoms to which they are attached form a homocyclic ring. substituted homocyclic ring, heterocyclic ring or substituted heterocyclic ring.

As used herein, the above terms have the following meaning:

“Alkyl” means a straight chain or branched, noncyclic or cyclic, unsaturated or saturated aliphatic hydrocarbon containing from 1 to 10 carbon atoms, while the term “lower alkyl” has the same meaning as alkyl but contains from 1 to 6 carbon atoms. The term “higher alkyl” has the same meaning as alkyl but contains from 2 to 10 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, and the like; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like. Representative saturated cyclic alkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like; while unsaturated cyclic alkyls include cyclopentenyl and cyclohexenyl, and the like. Cyclic alkyls are also referred to herein as a “homocycles” or “homocyclic rings.” Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as an “alkenyl” or “alkynyl”, respectively). Representative straight chain and branched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, and the like.

“Aryl” means an aromatic carbocyclic moiety such as phenyl or naphthyl.

“Arylalkyl” means an alkyl having at least one alkyl hydrogen atoms replaced with an aryl moiety, such as benzyl, —(CH₂)₂phenyl, —(CH₂)₃phenyl, —CH(phenyl)₂, and the like.

“Heteroaryl” means an aromatic heterocycle ring of 5- to 10 members and having at least one heteroatom selected from nitrogen, oxygen and sulfur, and containing at least 1 carbon atom, including both mono- and bicyclic ring systems. Representative heteroaryls are furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl.

“Heteroarylalkyl” means an alkyl having at least one alkyl hydrogen atom replaced with a heteroaryl moiety, such as —CH₂pyridinyl, —CH₂pyrimidinyl, and the like.

“Heterocycle” (also referred to herein as a “heterocyclic ring”) means a 4- to 7-membered monocyclic, or 7- to 10-membered bicyclic, heterocyclic ring which is either saturated, unsaturated, or aromatic, and which contains from 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, and wherein the nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized, including bicyclic rings in which any of the above heterocycles are fused to a benzene ring. The heterocycle may be attached via any heteroatom or carbon atom. Heterocycles include heteroaryls as defined above. Thus, in addition to the heteroaryls listed above, heterocycles also include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

“Heterocyclealkyl” means an alkyl having at least one alkyl hydrogen atom replaced with a heterocycle, such as —CH₂morpholinyl, and the like.

“Homocycle” (also referred to herein as “homocyclic ring”) means a saturated or unsaturated (but not aromatic) carbocyclic ring containing from 3-7 carbon atoms, such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclohexene, and the like.

The term “substituted” as used herein means any of the above groups (i.e., alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, homocycle, heterocycle and/or heterocyclealkyl) wherein at least one hydrogen atom is replaced with a substituent. In the case of a keto substituent (“—C(═O)—”) two hydrogen atoms are replaced. When substituted one or more of the above groups are substituted, “substituents” within the context of this invention include halogen, hydroxy, cyano, nitro, amino, alkylamino, dialkylamino, alkyl, alkoxy, alkylthio, haloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycle and heterocyclealkyl, as well as —NR_(a)R_(b), —NR_(a)C(═O)R_(b), —NR_(a)C(═O)NR_(a)NR_(b), —NR_(a)C(═O)OR_(b)—NR_(a)SO₂R_(b), —C(═O)R_(a), —C(═O)OR_(a), —C(═O)NR_(a)R_(b), —OC(═O)NR_(a)R_(b), —OR_(a), —SR_(a), —SOR_(a), —S(═O)₂R_(a), —OS(═O)₂R_(a) and —S(═O)₂OR_(a). In addition, the above substituents may be further substituted with one or more of the above substituents, such that the substituent substituted alky, substituted aryl, substituted arylalkyl, substituted heterocycle or substituted heterocyclealkyl. R_(a) and R_(b) in this context may be the same or different and independently hydrogen, alkyl, haloalkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl or substituted heterocyclealkyl.

“Halogen” means fluoro, chloro, bromo and iodo.

“Haloalkyl” means an alkyl having at least one hydrogen atom replaced with halogen, such as trifluoromethyl and the like.

“Alkoxy” means an alkyl moiety attached through an oxygen bridge (i.e. —O-alkyl) such as methoxy, ethoxy, and the like.

“Alkylthio” means an alkyl moiety attached through a sulfur bridge (i.e., —S-alkyl) such as methylthio, ethylthio, and the like.

“Alkylsulfonyl” means an alkyl moiety attached through a sulfonyl bridge (i.e. —SO₂-alkyl) such as methylsulfonyl, ethylsulfonyl, and the like.

“Alkylamino” and “dialkylamino” mean one or two alkyl moiety attached through a nitrogen bridge (i.e., —N-alkyl) such as methylamino, ethylamino, dimethylamino, diethylamino, and the like.

In one embodiment of this invention, A is O and representative GnRH receptor antagonists of this invention include compounds having the following structure (II):

In another embodiment, Q is —(CR_(8a)R_(8b))_(r)—Z—(CR_(10a)R_(10b))_(s)—, r and s are both zero, and representative GnRH receptor antagonists of this invention include compounds having the following structure (III):

In another embodiment, A is S, as represented by the following structure (IV):

Similarly, in another embodiment, A is NR₇, as represented by the following structure (V):

In further embodiments of this invention, R₆ is substituted or unsubstituted benzyl as represented by the following structure (VI) (wherein Y represents one or more optional substituents as defined above):

In a more specific embodiment of structure (VI), A is O, n is 2, and each occurrence of R_(3a) and R_(3b) is H, as represented by the following structure (VII):

With regard to the “R₁R₂N(CR_(3a)R_(3b))_(n)—” moiety of structure (I), n may be 2, 3 or 4. Accordingly, this moiety may be represented by the following structure (i) when n is 2, structure (ii) when n is 3, and structure (iii) when n is 3:

wherein each occurrence of R_(3a) and R_(3b) above may be the same or different, and are as defined above. For example, when each occurrence of R_(3a) and R_(3b) in structures (i), (ii) and (iii) is hydrogen, the “R₁R₂N(CR_(3a)R_(3b))_(n)—” moiety has the structure R₁R₂N(CH₂)₂—, R₁R₂N(CH₂)₃— and R₁R₂N(CH₂)₄—, respectively.

The compounds of the present invention may be prepared by known organic synthesis techniques, including the methods described in more detail in the Examples. However. in general, the compounds of structure (I) above may be made by the following Reaction Schemes. Specifically, compounds of structure (I) wherein A is oxygen may be made by Reaction Schemes A to E. Reaction Schemes F to K are appropriate for compounds of structure (I) wherein A is sulfur or NR₇, as well as where A is oxygen. Reaction Scheme L shows conditions for the conversion of thiouracils (where A is sulfur) to embodiments wherein A is NR₇. All substituents in the following Reaction Schemes are as defined above unless indicated otherwise.

Allylurea (i) and substituted acetoacetate (ii) are condensed under acidic conditions in a solvent such as ethanol or DMF at 25 to 100° C. and then cyclized under strongly basic conditions to give the substituted 3-allyl-2,4-pyrimidinedione (iii). Compound (iii) can then be modified by alkylation with an appropriate alkyl halide (where X is halogen) in a solvent such as DMF or ethanol for 1 hour to 2 days in the presence of a base such as sodium hydride or tetrabutylammonium fluoride to yield (iv). Oxidation of the allyl functionality, using osmium tetroxide and/or sodium periodate in solvent such as THF and/or water for 1-24 hours, gives aldehyde (v). Bromination of (v) using bromine or n-bromosuccinimide in a solvent such as acetic acid or chloroform for 1-24 hours resulted in brominated compound (vi). Reductive amination of (vi) with an appropriate amine using a reducing agent such as sodium triacetoxyborohydride in a solvent such as dichloroethane at 0 to 100° C. for 1-24 hours gives (vii) which when coupled with an appropriate boronic acid in a solvent such as ethanol or toluene at 25 to 150° C. for 1-24 hours in the presence of a Pd(0) catalyst gives (viii).

The final two steps of the above synthesis may also be reversed, the Suzuki coupling in that instance being the penultimate step and the reductive amination the final step. Alternatively, compound (iii) may be synthesized by the procedure in Example 2.

Compound (iii) from Reaction Scheme Al may also be synthesized by condensing and cyclizing allyl isocyanate (viii) and appropriate aminoalkene ester (ix) such as ethyl 3-aminocrotonate in a solvent such as toluene or DMF at 25 to 100° C. for 1-24 hours.

Cyclization of (xi) and (xii) in a solvent such as ethanol or DMF at 25 to 150° C. for 1 to 24 hours gives oxazime (xiii). Amination of (xiii) in a solvent such as DMF or ethanol at 25 to 150° C. for 1-24 hours yielded uracil derivative (xiv). Alkylation of (xiv) by an appropriate alkyl bromide in the presence of a base such as sodium hydride or sodium hydroxide in a solvent such as THF or DMF at 0 to 100° C. for 1-24 hours gives substituted uracil (xvi). The order of the reaction scheme may be changed allowing oxazine (xiii) to first be alkylated under conditions above to (xv) followed by amination to the product (xvi).

Compound (xvii) or (xviii) react with an appropriately substituted isocyanate in a solvent such as toluene or chloroform at room temperature to 100° C. for 1-24 hours as an alternative synthesis to intermediate oxazine (xv). Amination with a substituted amine in a solvent such as DMF or ethanol at a temperature of 25 to 100° C. for a period of 1-24 hours results in product uracil (xvi).

Intermediate (xvi) may be brominated using a brominating agent such as N-bromosuccinimide or bromine in a solvent such as acetic acid or chloroform at 0 to 100° C. for a period of 1-24 hours to yield bromo compound (ixx). The bromo compound can undergo various palladium catalyzed cross coupling reactions. Compound (ixx) taken in solvent such as ethanol or THF under nitrogen atmosphere using an appropriate Pd(0) catalyst such as tetrakis(triphenylphosphine)Pd(0), may be reacted for 1-24 hours at 25 to 150° C. with either an aryl boronic acid (ArB(OH)₂ where Ar is substituted aryl or heteroaryl) to yield product (xx) or with a substituted vinyl boronic acid to give compound (xxi). Compound (ixx) taken in solvent such as ethanol or THF using an appropriate Pd(0) catalyst in the presence of carbon monoxide and boronic acid yields (xxiv) after 1-24 hours at 0 to 150° C. Again using Pd(0) chemistry, compound (xxiii) is synthesized in a solvent such as THF or dioxane from the alkylation of (ixx) with an appropriate metal halide reagent for 1-24 hours at 0 to 150° C. Compound (ixx) in the presence of a substituted acetylene, Pd(0) catalyst, metal halide such as CuI, and base such as triethylamine in an appropriate solvent such as acetonitrile or DMF at 25 to 150° C. for 1-24 hours gives alkyne (xxii). Alkynyl uracil (xxii) may be selectively reduced to the alkene using a catalyst such as palladium/BaSO₄ under hydrogen atmosphere in solvent such as ethyl acetate or methanol to give (xxi).

Vinyl ester (xxvi) and (xxv) can be cyclized in a solvent such as DMF or EtOH at 25 to 150° C. for 1-24 hours to give (xxvii). Alkylation of (xxvii) with an appropriate alkyl or aryl halide in a solvent such as DMF or THF in the presence of a base such as sodium hydride or sodium hydroxide for 1-24 hours at 0 to 150° C. gives (xxviii).

Vinyl ester (xxvi) can be condensed with a substituted amine in a solvent such as DMF or ethanol at 25 to 150° C. for 1-24 hours to give (xxix). Cyclization of (xxix) with an isocyanate, isothiocyanate, or other appropriate compound in a solvent such as DMF, THF or dioxane, with or without a base such as sodium ethoxide or sodium hydride at 0 to 100° C. for 1-24 hours gives product (xxviii).

Compound (xxx) may be alkylated by an appropriate alkyl halide in the presence of a base such as sodium hydride or sodium hydroxide in a solvent such as THF or DMF at 0 to 50° C. for 1-24 hours to give (xxxi), which under further alkylation by a second alkyl halide gives product (xxviii).

Compound (xxxi) may be alkylated by an appropriate alkyl halide in the presence of a base such as sodium hydride or sodium hydroxide in a solvent such as THF or DMF at 0 to 100° C. for 1-24 hours to give (xxxii). The terminal double bond is oxidized using an appropriate oxidizing reagent such as osmium tetroxide or sodium periodate in solvent such as THF and/or water for 1-24 hours at 0 to 100° C. to give aldehyde (xxxiii). Reductive amination of (xxxiii) with an appropriate amine using a reducing agent such as sodium cyanoborohydride in a solvent such as dichloroethane or acetonitrile at 0 to 100° C. for 1-24 hours gives (xxviii).

Compound (xxxii) can be oxidized to the alcohol (xxxiv) first by hydroboration with a borane complex in a solvent such as THF followed by oxidation with ozone or hydrogen peroxide in a solvent such as methanol, ethanol and/or water at −25 to 100° C. for a period of 0.5-24 hours. Treatment of (xxxiv) with mesyl or tosyl chloride in methylene chloride with a base such as triethylamine or pyridine at 0 to 100° C. for 1-24 hours followed by reaction with an amine in a solvent such as DMF or toluene for 0.5-12 hours at 25 to 100° C. gives (xxviii).

Compound (xxxi) can be alkylated with an appropriate ester in a solvent such as DMF or ethanol in the presence of a base such as sodium hydride or sodium ethoxide at a temperature of 25 to 150° C. for a period of 1-24 hours to give (xxxv). Ester (xxxv) in a solvent such as chloroform or benzene with substituted amine and Lewis acid such as triethylaluminum gives amide (xxxvi) after 1-24 hours at 0 to 100° C. Reduction of (xxxvi) with lithium aluminum hydride or borane complex in a solvent such as THF or ether at 0 to 100° C. for 1-12 hours gives product (xxviii).

Thiouracil compound (xxxvii) in the presence of a substituted sulfonylisocyanate in a solvent such as benzene or toluene for 1-48 hours at 25 to 125° C. gives sulfonamide (xxxviii). Thiouracil (xxxvii) chlorinated by thionyl chloride or phosphorous oxychloride at −25 to 100° C. for 1-24 hours followed by amination with an appropriate amine in a solvent such as benzene or toluene at 25 to 150° C. for 1-24 hours gives compound (xxxix).

Substituted amine in the presence of urea or thiourea is heated at a temperature of 50-125° C. for 0.5 to 12 hours to give (xl). Cyclization of (xl) with diketene at 50-150° C. in acidic media such as acetic or formic acid for 5 minutes to 4 hours gives a mixture of isomers (xli) and (xlii). Halogenation of (xlii) using a halogenating reagent such as N-halosuccinimide in chloroform or bromine in acetic acid for 5 minutes to 24 hours gives halogenated product (xliii).

Uracil compound (xliii) and an appropriately substituted alcohol are condensed under Mitsonobu conditions such as diethyl or dibutyl axodicarboxylate and triphenylphosphine in a solvent such as THF at 0-100° C. for 0.5 to 10 hours to give compound (xliv). A Suzuki coupling of (xliv) and a boronic acid or boronic acid ester in a solvent such as ethanol or toluene at 25 to 150° C. for 1-24 hours in the presence of a Pd(0) catalyst gives (xlv). Deprotection of the protected amine gives (xlvi). Reductive amination of (xlvi) with an appropriate aldehyde in a solvent such as methylene chloride or acetonitrile using a reducing agent such as sodium triacetoxyborohydride or sodium borohydride at 0 to 100° C. for 1-24 hours gives (xlvii).

Keto or aldehyde xlviii in the presence of chlorosulfonylisocyanate or chlorocarbonylisocyanate yields oxaz-2,4-dione xlix after stirring for 1-24 hours at 0° C. to 75° C. in a solvent such as THF or ether. Mitsonobu condensation with an appropriate alcohol gives l which when in the presence of amine R₆NH₂ at room temperature to 125° C., with or without solvent such as DMF or catalyst such as acetic or hydrochloric acid, for ½ to 24 hours gives xlvii.

The compounds of the present invention may generally be utilized as the free acid or free base. Alternatively, the compounds of this invention may be used in the form of acid or base addition salts. Acid addition salts of the free amino compounds of the present invention may be prepared by methods well known in the art, and may be formed from organic and inorganic acids. Suitable organic acids include maleic, fumaric, benzoic, ascorbic, succinic, methanesulfonic, acetic, trifluoroacetic, oxalic, propionic, tartaric, salicylic, citric, gluconic, lactic, mandelic, cinnamic, aspartic, stearic, palmitic, glycolic, glutamic, and benzenesulfonic acids. Suitable inorganic acids include hydrochloric, hydrobromic, sulfuric, phosphoric, and nitric acids. Base addition salts included those salts that form with the carboxylate anion and include salts formed with organic and inorganic cations such as those chosen from the alkali and alkaline earth metals (for example, lithium, sodium, potassium, magnesium, barium and calcium), as well as the ammonium ion and substituted derivatives thereof (for example, dibenzylammonium, benzylammonium, 2-hydroxyethylammonium, and the like). Thus, the term “pharmaceutically acceptable salt” of structure (I) is intended to encompass any and all acceptable salt forms.

In addition, prodrugs are also included within the context of this invention. Prodrugs are any covalently bonded carriers that release a compound of structure (I) in vivo when such prodrug is administered to a patient. Prodrugs are generally prepared by modifying functional groups in a way such that the modification is cleaved, either by routine manipulation or in vivo, yielding the parent compound. Prodrugs include, for example, compounds of this invention wherein hydroxy, amine or sulfhydryl groups are bonded to any group that, when administered to a patient, cleaves to form the hydroxy, amine or sulfhydryl groups. Thus, representative examples of prodrugs include (but are not limited to) acetate, formate and benzoate derivatives of alcohol and amine functional groups of the compounds of structure (I). Further, in the case of a carboxylic acid (—COOH), esters may be employed, such as methyl esters, ethyl esters, and the like.

With regard to stereoisomers, the compounds of structure (I) may have chiral centers and may occur as racemates, racemic mixtures and as individual enantiomers or diastereomers. All such isomeric forms are included within the present invention, including mixtures thereof. Compounds of structure (I) may also possess axial chirality which may result in atropisomers. Furthermore, some of the crystalline forms of the compounds of structure (I) may exist as polymorphs, which are included in the present invention. In addition, some of the compounds of structure (I) may also form solvates with water or other organic solvents. Such solvates are similarly included within the scope of this invention.

The effectiveness of a compound as a GnRH receptor antagonist may be determined by various assay methods. Suitable GnRH antagonists of this invention are capable of inhibiting the specific binding of GnRH to its receptor and antagonizing activities associated with GnRH. For example, inhibition of GnRH stimulated LH release in immature rats may be measured according to the method of Vilchez-Martinez (Endocrinology 96:1130-1134, 1975). Briefly, twenty-five day old male Spraque-Dawley rats are administered an GnRH antagonist in saline or other suitable formulation by oral gavage, sub-cutaneous injection, or intravenous injection. This is followed by sub-cutaneous injection of 200 ng GnRH in 0.2 ml saline. Thirty minutes after the last injection, the animals are decapitated and trunk blood collected. After centrifugation, the separated plasma is stored at −20° C. until determination of the LH and FSH by radioimmunoassay. Other techniques for determining the activity of GnRH receptor antagonists are well known in the field, such as the use of cultured pituitary cells for measuring GnRH activity (Vale et al., Endocrinology 91:562-572, 1972), and a technique for measuring radioligand binding to rat pituitary membranes (Perrin et al., Mol. Pharmacol. 23:44-51, 1983).

For example, effectiveness of a compound as a GnRH receptor antagonist may be determined by one or more of the following assays.

Rat Anterior Pituitary Cell Culture Assay of GnRH Antagonists

Anterior pituitary glands are collected from 7-week-old female Sprague-Dawley rats and the harvested glands digested with collagenase in a dispersion flask for 1.5 hr at 37° C. After collagenase digestion, the glands are further digested with neuraminidase for 9 min at 37° C. The digested tissue is then washed with 0.1% BSA/McCoy's 5A medium, and the washed cells suspended in 3% FBS/0.1 BSA/McCoy's 5A medium and plated into 96-well tissue culture plates at a cell density of 40,000 cells per well in 200 μl medium. The cells are then incubated at 37° C. for 3 days. One pituitary gland normally yields one 96-well plate of cells, which can be used for assaying three compounds. For assay of an GnRH antagonist, the incubated cells are first washed with 0.1% BSA/McCoy's 5A medium once, followed by addition of the test sample plus 1 nM GnRH in 200 μl 0.1% BSA/McCoy's 5A medium in triplicate wells. Each sample is assayed at 5-dose levels to generate a dose-response curve for determination of its potency on the inhibition of GnRH stimulated LH and/or FSH release. After 4-hr incubation at 37° C., the medium is harvested and the level of LH and/or FSH secreted into the medium determined by RIA.

RIA of LH and FSH

For determination of the LH levels, each sample medium is assayed in duplicates and all dilutions are done with RIA buffer (0.01M sodium phosphate buffer/0.15M NaCl/1% BSA/0.01% NaN3, pH 7.5) and the assay kit is obtained from the Nation Hormone and Pituitary Program supported by NIDDK. To a 12×75 mm polyethylene test tube is added 100 μl of sample medium diluted 1:5 or rLH standard in RIA buffer and 100 μl of [125I]-labeled rLH (˜30,000 cpm) plus 100 μl of rabbit anti-rLH antibody diluted 1:187,500 and 100 μl RIA buffer. The mixture is incubated at room temperature over-night. In the next day, 100 μl of goat anti-rabbit IgG diluted 1:20 and 100 μl of normal rabbit serum diluted 1:1000 are added and the mixture incubated for another 3 hr at room temperature. The incubated tubes are then centrifuged at 3,000 rpm for 30 min and the supernatant removed by suction. The remaining pellet in the tubes is counted in a gamma-counter. RIA of FSH is done in a similar fashion as the assay for LH with substitution of the LH antibody by the FSH antibody diluted 1:30,000 and the labeled rLH by the labeled rFSH.

Radio-iodination of GnRH Peptide

The GnRH analog is labeled by the chloramine-T method. To 10 μg of peptide in 20 μl of 0.5M sodium phosphate buffer, pH 7.6, is added 1 mCi of Na125I, followed by 22.5 μg chloramine-T and the mixture vortexed for 20 sec. The reaction is stopped by the addition of 60 μg sodium metabisulfite and the free iodine is removed by passing the iodinated mixture through a C-8 Sep-Pak cartridge (Millipore Corp., Milford, Mass.). The peptide is eluted with a small volume of 80% acetonitrile/water. The recovered labeled peptide is further purified by reverse phase HPLC on a Vydac C-18 analytical column (The Separations Group, Hesperia, Calif.) on a Beckman 334 gradient HPLC system using a gradient of acetonitrile in 0.1% TFA. The purified radioactive peptide is stored in 0.1% BSA/20% acetonitrile/0.1% TFA at −80° C. and can be used for up to 4 weeks.

GnRH Receptor Membrane Binding Assay

Cells stably, or transiently, transfected with GnRH receptor expression vectors are harvested, resuspended in 5% sucrose and homogenized using a polytron homogenizer (2×15 sec). Nucleii are removed by centrifugation (3000×g for 5 min.), and the supernatant centrifuged (20,000×g for 30 min, 4° C.) to collect the membrane fraction. The final membrane preparation is resuspended in binding buffer (10 mM Hepes (pH 7.5), 150 mM NaCl, and 0.1% BSA) and stored at −70° C. Binding reactions are performed in a Millipore MultiScreen 96-well filtration plate assembly with polyethylenimine coated GF/C membranes. The reaction is initiated by adding membranes (40 ug protein in 130 ul binding buffer) to 50 ul of [¹²⁵I]-labeled GnRH peptide (˜100,000 cpm), and 20 ul of competitor at varying concentrations. The reaction is terminated after 90 minutes by application of vacuum and washing (2×) with phosphate buffered saline. Bound radioactivity is measured using 96-well scintillation counting (Packard Topcount) or by removing the filters from the plate and direct gamma counting. K_(i) values are calculated from competition binding data using non-linear least squares regression using the Prism software package (GraphPad Software).

Activity of GnRH receptor antagonists are typically calculated from the IC₅₀ as the concentration of a compound necessary to displace 50% of the radiolabeled ligand from the GnRH receptor, and is reported as a “K_(i)” value calculated by the following equation: $K_{i} = \frac{{IC}_{50}}{1 + {L/K_{D}}}$

where L=radioligand and K_(D)=affinity of radioligand for receptor (Cheng and Prusoff, Biochem. Pharmacol. 22:3099, 1973). GnRH receptor antagonists of this invention have a K_(i) of 100 μM or less. In a preferred embodiment of this invention, the GnRH receptor antagonists have a K_(i) of less than 10 μM, and more preferably less than 1 μM, and even more preferably less than 0.1 μM (i.e., 100 nM). To this end, representative GnRH receptor antagonists of this invention which have a K_(i) of less than 100 nM when using the GnRH receptor membrane binding assay as described above include the following Compound Nos.

Table No. Compound No. 1 3, 10, 11, 12, 13 3 1, 4 6 1, 2, 3, 8 7 2, 3, 4, 7, 9, 10, 11 8 2, 3, 4, 7, 12, 13, 14, 15, 16, 17, 19-21, 23, 25, 27-29, 31-36, 38-39, 42, 44, 51, 58, 59, 61, 63-66, 68, 70, 75, 77-97, 100, 106, 107, 109-113, 115-117, 124-135, 137-140 9 3, 4, 6, 7, 10, 14-16, 19, 24, 26, 32, 35, 37, 39, 40, 42, 46-49, 51-53, 55, 56, 58, 61, 63, 64, 66-68, 70, 72-78, 80-82, 85, 86, 89-93, 95, 96, 98-102, 107, 109, 110, 112, 138, 140, 142, 143, 145, 146, 149, 151-155, 157-162, 164, 166-168, 170-176, 178-188, 191, 194-197, 199, 200, 202-207, 210-212, 214, 215, 219, 224, 225, 227, 229, 232-234, 237, 240, 242, 244, 245, 247, 249, 251-256, 258-261, 263, 265-267, 270, 275, 277-279, 281, 286, 287, 295-301, 304, 305, 307-309, 312, 318, 320, 321, 325-329, 331-336, 338-346, 348-355, 357-359, 361, 362, 364-385, 387-397, 399, 402, 406, 409, 410, 413, 415, 417, 419-424, 427-434, 437-439, 441, 443, 446, 448, 454, 455, 470, 473, 477, 480-487, 490-493, 495, 502, 503, 509, 512, 514, 517, 519-524, 547-552, 554-560, 565-568, 570, 581-584, 589, 595, 596, 602, 606-609, 612, 613, 618, 621, 622, 624-627, 634, 636, 642-648, 652, 653, 655-658, 660-662, 664, 665, 668-672, 677, 678, 680, 681, 688, 694, 696, 698-702, 704, 706-708, 711, 712, 714, 718-726, 729-741, 745, 747-750, 755-756, 759-763, 774 10 1, 10, 14, 21-23, 25, 52, 54-56, 60, 61, 64, 65 12 1, 4, 5, 10, 20-22, 24, 27, 32 13 2, 4 15 1, 2

As mentioned above, the GnRH receptor antagonists of this invention have utility over a wide range of therapeutic applications, and may be used to treat a variety of sex-hormone related conditions in both men and women, as well as mammals in general. For example, such conditions include endometriosis, uterine fibroids, polycystic ovarian disease, hirsutism, precocious puberty, gonadal steroid-dependent neoplasia such as cancers of the prostate, breast and ovary, gonadotrophe pituitary adenomas, sleep apnea, irritable bowel syndrome, premenstrual syndrome, benign prostatic hypertrophy, contraception and infertility (e.g., assisted reproductive therapy such as in vitro fertilization).

The compounds of this invention are also useful as an adjunct to treatment of growth hormone deficiency and short stature, and for the treatment of systemic lupus erythematosis.

In addition, the compounds are useful in combination with androgens, estrogens, progesterones, and antiestrogens and antiprogestogens for the treatment of endometriosis, fibroids, and in contraception, as well as in combination with an angiotensin-converting enzyme inhibitor, an angiotensin II-receptor antagonist, or a renin inhibitor for the treatment of uterine fibroids. The compounds may also be used in combination with bisphosphonates and other agents for the treatment and/or prevention of disturbances of calcium, phosphate and bone metabolism, and in combination with estrogens, progesterones and/or androgens for the prevention or treatment of bone loss or hypogonadal symptoms such as hot flashes during therapy with a GnRH antagonist.

In another embodiment of the invention, pharmaceutical compositions containing one or more GnRH receptor antagonists are disclosed. For the purposes of administration, the compounds of the present invention may be formulated as pharmaceutical compositions. Pharmaceutical compositions of the present invention comprise a GnRH receptor antagonist of the present invention and a pharmaceutically acceptable carrier and/or diluent. The GnRH receptor antagonist is present in the composition in an amount which is effective to treat a particular disorder—that is, in an amount sufficient to achieve GnRH receptor antagonist activity, and preferably with acceptable toxicity to the patient. Typically, the pharmaceutical compositions of the present invention may include a GnRH receptor antagonist in an amount from 0.1 mg to 250 mg per dosage depending upon the route of administration, and more typically from 1 mg to 60 mg. Appropriate concentrations and dosages can be readily determined by one skilled in the art.

Pharmaceutically acceptable carrier and/or diluents are familiar to those skilled in the art. For compositions formulated as liquid solutions, acceptable carriers and/or diluents include saline and sterile water, and may optionally include antioxidants, buffers, bacteriostats and other common additives. The compositions can also be formulated as pills, capsules, granules, or tablets which contain, in addition to a GnRH receptor antagonist, diluents, dispersing and surface active agents, binders, and lubricants. One skilled in this art may further formulate the GnRH receptor antagonist in an appropriate manner, and in accordance with accepted practices, such as those disclosed in Remington's Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co., Easton, Pa. 1990.

In another embodiment, the present invention provides a method for treating sex-hormone related conditions as discussed above. Such methods include administering of a compound of the present invention to a warm-blooded animal in an amount sufficient to treat the an condition. In this context, “treat” includes prophylactic administration. Such methods include systemic administration of a GnRH receptor antagonist of this invention, preferably in the form of a pharmaceutical composition as discussed above. As used herein, systemic administration includes oral and parenteral methods of administration. For oral administration, suitable pharmaceutical compositions of GnRH receptor antagonists include powders, granules, pills, tablets, and capsules as well as liquids, syrups, suspensions, and emulsions. These compositions may also include flavorants, preservatives, suspending, thickening and emulsifying agents, and other pharmaceutically acceptable additives. For parental administration, the compounds of the present invention can be prepared in aqueous injection solutions which may contain, in addition to the GnRH receptor antagonist, buffers, antioxidants, bacteriostats, and other additives commonly employed in such solutions.

The following example is provided for purposes of illustration, not limitation. In summary, the GnRH receptor antagonists of this invention may be assayed by the general methods disclosed above, while the following Examples disclose the synthesis of representative compounds of this invention.

EXAMPLE 1 SYNTHESIS OF 1-(2,6-DIFLUOROBENZYL)-5-(3-METHOXYPHENYL)-6-METHYL-3-[N-METHYL-N-(2-PYRIDYLETHYL)AMINOETHYL]URACIL

Step 1A 3-Allyl-6-methyluracil

To allylurea (25 g, 0.25 mol) in ethanol (10 mL) was added ethyl acetoacetate (31.86 mL, 0.25 mol) and 10 drops conc. HCl. After 12 days at room temperature, concentration gave an oil which was dissolved in MeOH. KOH (22.5 g, 0.34 mol) was added and the solution refluxed for 1 hour. After neutralization, the resulting solid 1 was collected. Yield 2.7 g (7%). NMR (CDCl₃) δ: 2.16 (3H, s), 4.52 (2H, d), 5.18 (1H, d), 5.23 (1H, d), 5.60 (1H, s), 5.82-5.93 (1H, m), 10.3 (1H, s).

Step 1B 3-Allyl-1-(2,6-difluorobenzyl)-6-methyluracil

To 1 (2.6 g, 15.7 mmol) in DMF (20 mL) was added tetrabutylammoniumfluoride (25 mmol) and 2,6-difluorobenzyl bromide (4.14 g, 20 mmol). After 2 days stirring at room temperature, column chromatography using ethyl acetate/hexane gave 2.7 g (59% yield) of 2. MS 293 (MH)⁺.

Step 1C 3-Acetaldehyde-1-(2,6-difluorobenzyl)-6-methyluracil

To a solution of 2 (1.46 g, 5 mmol) in THF (20 mL) and H₂O (10 mL) was added osmium tetroxide (200 mg) and NaIO₄ (3.2 g, 15 mmol). After 2 hr, another 1 g of NaIO₄ was added. Ethyl acetate and H₂O were added and the layers separated. Evaporation of the organic layer gave 3 as a crude solid (1.0 g, 68%). MS 295 (MH)⁺.

Step 1D 3-Acetaldehyde-5-bromo-1-(2,6-difluorobenzyl)-6-methyluracil

3 (294 mg, 1 mmol) was dissolved in acetic acid and bromine (1.2 eq) was added. The reaction mixture was stirred at room temperature for 1 hr, evaporated and the residue was dissolved in EtOAc, washed with 1N KOH solution and concentrated to give 4 as a crude oil (295 mg, 79%). MS 373/375 (MH)⁺. NMR (CDCl₃) δ: 2.55 (3H, s), 4.87 (2H, d), 5.33 (2H, s), 7.26-7.33 (3H, 2m), 9.59 (1h, d).

Step 1E 5-Bromo-1-(2,6-difluorobenzyl)-6-methyl-3-[N-methyl-N-(2-pyridylethyl)aminoethyl]uracil

To 4 (295 mg, 0.8 mmol) in dichloroethane was added 2-(methylaminoethyl)pyridine (200 mg, 1.5 mmol) and NaBH(OAc)₃ (636 mg, 3 mmol). After overnight stirring, the reaction mixture was concentrated, dissolved in EtOAc, washed with H₂O, and purified by prep TLC to give 190 mg of 5 (48%).

Step 1F 1-(2,6-Difluorobenzyl)-5-(3-methoxyphenyl)-6-methyl-3-[N-methyl-N-(2-pyridylethyl)aminoethyl]uracil (“Cpd. No. 1”)

5 (150 mg, 0.3 mmol), 3-methoxyphenylboronic acid (92 mg, 0.6 mmol), K₂CO₃ (100 mg, 0.72 mmol), and Pd(PPh₃)₄ (20 mg) in H₂O (5 mL ) and toluene (10 mL) was heated a sealed tube at 100° C. for 12 hr. Purification by HPLC gave 40 mg of 6 (“Cpd. No. 1”) as the TFA salt (21% yield). MS 521 (MH)⁺ NMR (CDCl₃) δ: 2.14 (3H, s), 3.02 (3H, s), 3.50 (2H, m), 3.63 (2H, m), 3.71 (2H, m), 3.81 (3H, s), 4.37 (2H, m), 5.25 (2H, s), 6.81-6.83 (2H, m), 6.88-6.95 (3H, m), 7.28-7.34 (2H, m), 7.63 (1H, m), 7.89 (1H, d), 8.13 (1H, t), 8.62 (1H, br s).

EXAMPLE 2 REPRESENTATIVE COMPOUNDS

Following the procedures as set forth in Example 1 above, the compounds of the following Table 1 were prepared.

TABLE 1

Cpd. No. R₁ R₂ MS (MH)⁺ 1-1  2-PyCH₂CH₂ H 507 1-2  2-PyCH₂ H 493 1-3  2-PyCH₂ Me 507 1-4  Bz Me 506 1-5  PhCH₂CH₂ Me 520 1-6  2-PyCH₂CH₂ Pr 549 1-7  PhCHCH₃ Me 520 1-8  PhCHCH₃ Me 520 1-9  Bz (CH₃)₂N(CH₂)₂ 563 1-10 2-PyCH₂CH₂ Et 535 1-11 2-(6-Cl-Py)CH₂CH₂ Me 416, 555 1-12 2-PyCH₂CH₂ CyclopropylCH₂ 561 1-13 1-Et-3-pyrrolidinyl Et 527 1-14

H 557 1-15

H 541 1-16 (CH₃)₂CHOCH₂CH₂CH₂ H 502 1-17 Et₂NCH₂CH₂ Me 515 1-18

H 513 1-19 CH₃OCH₂CH₂CH₂ H 474 1-20 (EtO)₂CHCH₂CH₂ H 532 1-21 CH₃OCH₂CH₂ Me 474 1-22

Me 575 1-23

Me 575 1-24

H 550 1-25 CH₃OCH₂CH₂CH₂ Me 488 1-26 (EtO)₂CHCH₂CH₂ Me 546 1-27

Me 564 1-28

Me 571 1-29

Me 555 1-30 (CH₃)₂CHOCH₂CH₂CH₂ Me 516 1-31

Me 527 1-32

Me 513 1-33

Me 502 1-34 Et₂NCH₂CH₂ Me 487 1-35 Me₂NCH₂CH₂CH₂ Me 501 1-36 Et₂NCH₂CH₂CH₂ Me 529 1-37

Me 499 1-38 EtOCH₂ Me 474 1-39

Me 516 1-40

Me 550 1-41

H 513 1-42

H 496 1-43

H 529 1-44 Me₂NCH₂CH₂CH₂ H 487 1-45 Et₂NCH₂CH₂CH₂ H 515 1-46

H 510 1-47

H 541 1-48 Me₂CHCH₂OCH₂CH₂CH₂ H 516 1-49

H 502 1-50

H 471 1-51

H 471 1-52

H 536 1-53

H 516 1-54 PyCH₂CH₂ HOCH₂CH₂ 551 1-55

Me 527 1-56

Me 510 1-57

Me 543 1-58 Me₂CHN(Me)CH₂CH₂CH₂ Me 529 1-59

Me 524 1-60

Me 555 1-61 Me₂CHCH₂OCH₂CH₂CH₂ Me 530 1-62 BuOCH₂CH₂CH₂ Me 530 1-63

Me 499 1-64

Me 499 1-65

Me 550 1-66

Me 530 1-67 PhCH₂CH₂CH₂ H 506 1-68

Me 535

EXAMPLES 3 Further Representative Compounds

By reversing Step 1E and Step 1F in Example 1, where the boronic acid coupling is performed followed by the reductive amination, the compounds of the following Tables 2-7 were also prepared.

TABLE 2

Cpd. No. R₁ R₂ MS (MH)⁺ 2-1 2-PyCH₂CH₂ Me 519 2-2 Bz Me 504 2-3 2-PyCH₂ H 491 2-4 2-PyCH₂CH₂ H 505 2-5 PhCH₂CH₂ Me 518

TABLE 3

Cpd. No. R₁ R₂ MS (MH)⁺ 3-1 2-PyCH₂CH₂ Me 535 3-2 PhCH₂CH₂ Me 534 3-3 4-PyCH₂CH₂ Me 535 3-4 2-PyCH₂CH₂ Et 549

TABLE 4

Cpd. No. R₁ R₂ MS (MH)⁺ 4-1 PhCH₂ Me 474 4-2 2-PyCH₂CH₂ Me 489 4-3

518 4-4

520 4-5

491 4-6

547

TABLE 5

Cpd. No. R₁ R₂ MS (MH)⁺ 5-1 PhCH₂CH₂ Me 488 5-2 2-PyCH₂CH₂ Me 503 5-3

545 5-4

559

TABLE 6

Cpd. No. R₄ MS (MH)⁺ 6-1 

509 6-2 

583 6-3 

549 6-4 

481 6-5 

551 6-6 

495 6-7 

519 6-8 

606 6-9 

497 6-10

525 6-11

559/561 6-12

525 6-13

519 6-14

505 6-15

551 6-16

548 6-17

490 6-18

504 6-19

516 6-20

575 6-21

543 6-22

535 6-23

581 6-24

541 6-25

575 6-26

521 6-27

519 6-28

531 6-29

533 6-30

567 6-31

519 6-32

537 6-33

521 6-34

581 6-35

509 6-36

509 6-37

536 6-38

497 6-39

535 6-40

519 6-41

567 6-42

481 6-43

497 6-44

507 6-45

548 6-46

573 6-47

584 6-48

645 6-49

534 6-50

535 6-51

506 6-52

562 6-53

533 6-54

516 6-55

505

TABLE 7

Cpd. No. NR₁R₂ MS (MH)⁺ 7-1 

486 7-2 

539 7-3 

567 7-4 

571 7-5 

590 7-6 

527 7-7 

486 7-8 

514 7-9 

530 7-10

484 7-11

513 7-12

546 7-13

553 7-14

485 7-15

485 7-16

499 7-17

499 7-18

513 7-19

472 7-20

546

EXAMPLE 4 SYNTHESIS OF 5-BROMO-1-(2,6-DIFLUOROBENZYL)-6-METHYL-URACIL

Step A 2,6-Difluorobenzyl Urea

2,6-Difluorobenzylamine (25.0 g, 0.175 mol) was added dropwise to a stirring solution of urea (41.92 g, 0.699 mol) in water (70 mL) and concentrated HCl (20.3 mL). The resulting mixture was refluxed for 2.5 hours, after which time it was cooled to room temperature. The solids that formed were filtered under vacuum, and were washed thoroughly with water. After drying under vacuum, the solids were recrystallized from EtOAc to yield the product 1 as light white needles (24.0 g, 0.129 mol, 74%).

Step B 1-(2,6-Difluorobenzyl)-6-methyl-uracil

Diketene (9.33 mL, 0.121 mol) was added in one portion to a refluxing solution of 2,6-difluorobenzyl urea 1 (20.46 g, 0.110 mol) and glacial acetic acid (110 mL). After 40 minutes at reflux, the mixture was cooled to room temperature and poured onto water (600 mL). The precipitate was collected by filtration, washed with water and dried under vacuum to yield a 1:3 mixture of isomers 2 and 3, respectively (19.07 g, 0.076 mol, 69%). The mixture was recrystallized from acetonitrile (˜600 mL) to give the pure title compound 3 as white prisms (1^(st) crop—7.85 g, 0.031 mol, 28%).

Step C 5-Bromo-1-(2,6-difluorobenzyl)-6-methyl-uracil

1-(2,6-Difluorobenzyl)-6-methyl-uracil 3 (7.56 g, 30 mmol) was suspended in glacial acetic acid (100 mL) and to that mixture, bromine (1.93 mL, 37.5 mmol) was added dropwise. The resulting orange solution turned into a suspension in about 5 minutes. After stirring for 1 hour at room temperature, the precipitate was filtered under vacuum and washed with water. The solids were triturated with diethyl ether and dried under vacuum to give 4 (8.6 g, 0.026 mmol, 87%).

EXAMPLE 5 FURTHER REPRESENTATIVE COMPOUNDS

Step A-1 3-(1-[2-BOC-(S)-amino-3-phenylpropyl)-5-bromo-1-(2,6-difluorobenzyl)-6-methyl-uracil

2-BOC-(S)-amino-3-phenyl-1-propanol (2.51 g, 10 mmol) and triphenylphosphine (3.14 g, 12 mmol) were added to a solution of 5-bromo-1-(2,6-difluorobenzyl)-6-methyl-uracil 1 (3.31 g, 10 mmol) in THF (50 ml). Di-tert-butyl azodicarboxylate (2.76 g, 12 mmol) was added in several portions over 5 minutes. After 5 minutes the reaction mixture was clear. After 1 hour the reaction mixture was concentrated and the residue was purified by silica cartridge column (hexane/EtOAc as elutant). Concentration of like fractions gave 6.8 g of an oily material which was precipitated from hexane to yield product 2 (4.95 g, 88%).

Step B-1 3-(1-[2-BOC-(S)-amino-3-phenylpropyl)-1-(2,6-difluorobenzyl)-5-(2-fluoro-3-methoxyphenyl)-6-methyl-uracil

Compound 2 (4.95 g, 8.78 mmol) and sodium carbonate (2.12g, 20 mmol) were suspended in toluene (50 mL) and dimethoxyethane (10 mL). Water (20 mL) was added and N₂ was bubbled through the reaction mixture. After 5 minutes, both layers were clear and Pd(OAc)₂ (394 mg, 0.2 eq) and triphenylphosphine ((921 mg, 0.4 eq) were added. The boronic acid (1.7 g, 10 mmol) was added and the reaction vessel was sealed and heated overnight at 100° C. The organic layer was separated, evaporated and purified by silica chromatography. Product containing fractions were combined and evaporated to give 3 as a brown oil (1.5 g, 28% yield).

Step C-1 3-(1-[2-(S)-Amino-3-phenylpropyl)-1-(2,6-difluorobenzyl)-5-(2-fluoro-3-methoxyphenyl)-6-methyl-uracil

Compound 3 (1.5 g, 2.5 mmol) in trifluoroacetic acid/dichloromethane (1:1, 50 mL) was heated for 4 hours. Evaporation gave a red oil which was purified by reverse phase prep HPLC using water/CH₃CN with 0.05% trifluoroacetic acid as elutant. The product containing fractions were concentrated and lyophilized to give product 4 (0.56 g, 44%, MH⁺=510).

Step A-2 1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-6-methyl-5-(4-[tetrahydropyran-2-yloxy]phenyl]uracil

1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-6-methyl-5-bromouracil 1 (2.58 g, 4.7 mmol), tetrakis(triphenylphosphine) palladium (0) (550 mg, 0.47 mmmol), 4-hydroxyphenyl boronic acid tetrahydropyran ether (1.25 g, 5.7 mmol) and barium hydroxide (38 mL of 0.14M solution, 5.2 mmol) in a benzene/ethanol/dimethoxyethane solution (10/1/11, 90 mL) was heated at 90° C. in a pressure vessel under N₂ atmosphere overnight. The organic layer was concentrated in vacuo and the residue was purified by silica gel chromatography (hexanes/ethyl acetate as elutant) to give 3.0 g of 2 as an off white foam.

Step B-2 1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-6-methyl-5-(4-hydroxyphenyl)uracil

A mixture of 2 (3.0 g, 4.6 mmol) and pyridinium-p-toluenesulfonate (231 mg, 0.92 mmol) in ethanol (92 mL) was stirred at 45° C. for 5 hours. The reaction mixture was concentrated in vacuo and the residue was dissolved in methylene chloride and H₂O. The organic layer was concentrated and the residue purified by silica gel chromatography using hexanes/ethyl acetate as elutant to give 2.1 g of compound 3 as a yellow foam.

Step C-2 1-(2,6-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(4-[4-tolyloxy]phenyl)uracil

Substituted uracil 3 (50 mg, 0.089 mmol), p-tolylboronic acid (18 mg, 0.133 mmol), copper (II) acetate (16 mg, 0.089 mmol) and triethylamine (0.06 mL, 0.445 mmol) in CH₂Cl₂ (1 mL) were stirred for 3 days at room temperature. The reaction mixture was purified by silica gel chromatography using 1% MeOH in CH₂Cl₂ to give 30 mg of protected product. This material was dissolved in CH₂Cl₂ (1 mL) with 5 drops of trifluoroacetic acid. Purification by reverse phase HPLC/MS gave 5.0 mg of product 4 m/z (CI) 554 (MH⁺).

Step A-3 (S)-3-(1-N-tert-Butoxycarbonylamino-1-carboxylic acid ethyl)-1-(2,6-difluorobenzyl)-5-(3-methoxyphenyl)-6-methyluracil

To a stirred solution of 1 (306 mg, 0.55 mmol) in tetrahydrofuran (15 mL) at room temperature, was added aqueous lithium hydroxide solution (15 mL of a 1 M solution, 15 mmol). After 2 h, most of the tetrahydrofuran was removed in vacuo and the resulting solution was acidified to pH 4 (with 10% aqueous citric acid solution). The resultant precipitate was extracted into ethyl acetate (2×15 mL) and the combined organic layer was washed with water, brine and dried (MgSO₄). The solvent was removed in vacuo to give 2 (283 mg, 94%) as a yellow oil which was not purified further, d_(H) (300 MHz; CDCl₃) 7.26-7.34 (2H, m, Ar), 6.73-6.95 (5H, m, Ar), 5.74(1H, brd, J 6, NH), 5.37(1H, d, J 16, CHHAr), 5.22 (1H, d, J 16, CHHAr), 4.62 (1) brs, CHN), 4.32-4.49 (2H, m, CH₂N), 3.80 (3H, s, OCH₃), 2.17 (3H, s, CH₃) and 1.42 (9H, s, 3×CH₃), m/z (CI) 446 (MH⁺-Boc, 100%).

Step B-3 (S)-3-(1-Amino-1-NH-benzylcarboxamide ethyl)-1-(2,6-difluorobenzyl)-5-(3-methoxyphenyl)-6-methyluracil Trifluoroacetic Acid Salt

To a stirred solution of 2 (20 mg, 0.037 mmol), benzylamine (15 μL, 0.14 mmol), 1-(hydroxy)benzotriazole hydrate (9 mg, 0.066 mmol) and triethylamine (10 μL, 0.074 mmol) in anhydrous N,N-dimethylformamide (1 mL) at room temperature, was added 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (11 mg, 0.056 mmol). After 10 h, the reaction mixture was poured into water (ca. 5 mL) and the resulting precipitate was extracted into ethyl acetate (ca. 5 mL). The organic layer was washed with brine and dried (MgSO₄). The solvent was removed in vacuo to give a yellow oil, which was redissolved in a mixture of dichloromethane (1 mL) and trifluoroacetic acid (0.5 mL, 6.5 mmol) and stirred at room temperature. After 1 h, the solvent was removed in vacuo to give a yellow oil, which was purified by reverse phase HPLC/MS to give 3 (6 mg, 30%) as a colorless solid, m/z (CI) 535.2 (MH⁺, 100%).

By the above procedure, the compounds of the following Table 8 were also prepared.

TABLE 8

MS MS Cpd. No. R₁R₂N(CR_(3a)R_(3b))_(n)— —Q—R₄ (calc) Ion 8-1

491.5 492 8-2

509.5 510 8-3

509.5 510 8-4

H 385.4 386 8-5

415.4 416 8-6

433.4 434 8-7

H 385.4 386 8-8

415.4 416 8-9

415.4 416 8-10

433.4 434 8-11

515.6 516 8-12

H 391.5 392 8-13

495.5 496 8-14

495.5 496 8-15

539.6 540 8-16

477.5 478 8-17

479.5 480 8-18

539.6 540 8-19

505.5 506 8-20

501.5 502.2 8-21

501.5 502.2 8-22

465.5 450 8-23

475.5 476.2 8-24

461.5 462.2 8-25

461.5 462.2 8-26

487.5 488.2 8-27

487.5 488.2 8-27

487.5 488.2 8-28

527.5 528 8-29

523.6 524 8-30

523.6 524 8-31

495.5 496 8-32

475.5 476.2 8-33

491.5 492 8-34

539.6 540 8-35

481.9 482 8-36

493.6 494 8-37

525.6 526 8-38

489.5 490.2 8-39

475.5 476.2 8-40

449.4 450 8-41

445.4 446 8-42

467.5 469 8-43

449.5 450 8-44

479.5 480 8-45

459.4 460.2 8-46

445.4 446.1 8-47

534.6 535.2 8-48

514.5 515.2 8-49

521.5 522.2 8-50

548.6 549.2 8-51

520.3 521.2 8-52

512.6 513.2 8-53

527.6 528.2 8-54

502.5 503.2 8-55

528.6 529.3 8-56

H 371.4 372.1 8-57

490.6 491.2 8-58

478.5 479.1 8-59

491.5 492.2 8-60

515.5 516.2 8-61

493.6 494.1 8-62

553.6 554.2 8-63

553.6 554.2 8-64

553.6 554.2 8-65

557.6 558.2 8-66

557.6 558.2 8-67

569.6 570.2 8-68

569.6 570.2 8-69

574.0 574.2 8-70

574 574.2 8-71

581.7 582.3 8-72

595.7 596.3 8-73

607.6 608.2 8-74

608.5 608.1 8-75

488 488.1 8-76

489.5 490.2 8-77

447.5 488.2 8-78

465.5 466.1 8-79

513.5 514.1 8-80

495.5 496.2 8-81

509.5 510.1 8-82

465.5 466.2 8-83

495.5 496.2 8-84

513.5 514.2 8-85

509.5 510.2 8-86

461.5 462 8-87

477.5 478 8-88

481.9 482 8-89

461.5 462 8-90

515.5 516 8-91

489.6 490 8-92

531.5 532 8-93

523.6 524 8-94

465.5 466 8-95

481.9 482 8-96

461.5 462 8-97

475.5 476 8-98

503.6 504 8-99

523.6 524 8-100

477.5 478 8-101

531.5 532 8-102

491.5 482 8-103

497.5 498 8-104

516.4 516 8-105

475.5 476 8-106

467.5 468 8-107

453.5 454 8-108

476.5 474 8-109

489.6 490 8-110

465.5 466.1 8-111

495.5 496.2 8-112

513.5 514.2 8-113

509.5 510.1 8-114

498.6 498 8-115

545.5 546.2 8-116

563.5 564.2 8-117

559.5 560.2 8-118

561.5 562.2 8-119

579.5 580.2 8-120

575.5 576.2 8-121

481.9 482.1 8-122

525.9 526.1 8-123

512 512.1 8-124

529.9 530.1 8-125

483.5 484.1 8-126

513.5 496.2 8-127

531.5 532.1 8-128

491.5 492.2 8-129

513.5 514.2 8-130

527.5 528.1 8-131

531.5 532.2 8-132

483.5 484.1 8-133

513.5 514.2 8-134

527.5 528.2 8-135

531.5 532.2 8-136

483.5 484.1 8-137

513.5 514.1 8-138

527.5 528.2 8-139

531.5 532.2 8-140

483.5 484.1

EXAMPLE 6 Synthesis of Boronic Acids

Step A 2-Fluoro-3-methoxyphenylboronic Acid

n-Butyl lithium (20 mL, 2.5M) was added to a solution of tetramethylpiperidine (8.44 mL, 50 mmol) in THF (125 mL) at −78° C. The reaction mixture was stirred at −78° C. for 1.5 hours. 2-Fluoroanisole (6.31 g, 50 mmol) was added and the mixture was stirred for 8 hours at −78° C. Trimethyl borate (6.17 mL, 55 mmol) was added and the reaction mixture was allowed to warm slowly to room temperature overnight. The mixture was poured into 1N HCl (250 mL). Extraction with EtOAc followed by evaporation gave a sticky solid which was triturated with hexanes to give product (2.19 g, 26% yield).

EXAMPLE 7 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A BOC-(S)-1-amino-2-propanol

Di-t-butyl dicarbonate (6.76 g, 31 mmol) was added portionwise to a stirred solution of(S)-1-amino-2-propanol and triethylamine (4.4 mL, 31.5 mmol) in CH₂Cl₂ (75 mL) at 0° C. The reaction mixture was stirred for 1 hour at 0° C. and for 30 minutes at room temperature. Evaporation gave product 1 which was used without further purification.

Step B 3-(2-BOC-(R)-1-aminopropyl)-5-bromo-1-(2,6-difluorobenzyl)-6-methyl-uracil

5-Bromo-1-(2,6-difluorobenzyl)-6-methyluracil (3.31 g, 10 mmol) was suspended in THF (200 mL). Compound 1 (1.84 g, 10.5 mmol) and triphenylphosphine (3.93 g, 15 mmol) were added and the mixture was stirred. DEAD (2.36 mL, 15 mmol) was added and the reaction mixture became a solution. After stirring overnight, the volatiles were removed and the residue was chromatographed on silica using EtOAc/hexanes as elutant to give white solid 2 (4.57 g, 94% yield).

EXAMPLE 8 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A

A solution of N-(t-butyloxycarbonyl)-D-α-alaninol (1.75 g, 10 mmol) in anhydrous THF (15 mL) was treated with 5-bromo-1-(2,6-difluorobenzyl)-6-methyluracil (3.31 g, 10 mmol) and triphenylphosphine (3.15 g, 12 mmol) at ambient temperature, then di-tert-butylazodicarboxylate (2.76 g, 12 mmol) was introduced. The reaction mixture was stirred at ambient temperature for 16 hours and volatiles were evaporated. The residue was partitioned between saturated NaHCO₃/H₂O and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by flash chromatography (silica, 1:2 EtOAc/hexanes) to give compound 1 (4.69 g, 96.1%), MS (CI) m/z 388.0, 390.0 (MH⁺-Boc).

Step B

To compound 1 (1.0 g, 2.05 mmol) in benzene/EtOH/ethylene glycol dimethyl ether (20/2/22 mL) was added 2-fluoro-3-methoxyphenylboronic acid (435 mg, 2.56 mmol) and saturated Ba(OH)₂/water (˜0.5 M, 15 mL). The reaction mixture was deoxygenated with N₂ for 10 min. tetrakis(triphenylphosphine)palladium (0) (242 mg, 0.21 mmol) was added and the reaction mixture was heated at 80° C. overnight under the protection of N₂. The reaction mixture was partitioned between brine and EtOAc. The organic layer was dried (sodium sulfate), evaporated, purified by flash chromatography (silica, 40% EtOAc/hexanes) to give compound 2 (450 mg, 41.2%), MS (CI) m/z 434.2 (MH⁺-Boc).

Step C

TFA (2 mL) was added to a solution of 2 (267 mg, 0.5 mmol) in dichloromethane (2 mL) and the reaction mixture was stirred at ambient temperature for 1 hour. Volatiles were evaporated and the residue was partitioned between saturated NaHCO₃/water and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by reverse phase HPLC (C-18 column, 15-75% acetonitrile/water) to give compound 3, MS (CI) m/z 434.2 (MH⁺).

Step D

2-Pyridinecarboxyaldehyde (80 mg, 0.75 mmol) was added to a solution of 3 (267 mg, 0.5 mmol) in MeOH (5 mL) and the reaction mixture was stirred at ambient temperature for 10 hours. NaBH₄ (56 mg, 1.5 mmol) was added and the reaction mixture was kept at ambient temperature for 10 minutes. Volatiles were evaporated and the residue was partitioned between saturated NaHCO₃/water and dichloromethane. The organic layer was dried (sodium sulfate), evaporated, and purified by reverse phase HPLC (C-18 column, 15-75% acetonitrile/water) to give compound 4, MS (CI) m/z 525.20 (MH⁺).

Step E

To a solution of 4 (20 mg, 0.04 mmol) in dichloroethane (2 mL) was added 1 drop of formaldehyde (37% solution in water) and NaBH(OAc)₃ (16 mg, 0.08 mmol). The reaction mixture was stirred at ambient temperature for 2 hours, volatiles were evaporated and the residue was partitioned between water and dichloromethane. The organic layer was dried (sodium sulfate), evaporated, and purified by reverse phase HPLC (C-18 column, 15-75% acetonitrile/water) to give compound 5, MS (Cl) m/z 539.20 (MH⁺).

EXAMPLE 9 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A

A solution of N^(α)-(t-butyloxycarbonyl)-L-α-cyclohexylglycine (2.0 g, 7.77 mmol) in anhydrous THF (10 mL) was cooled down to 0° C. Borane solution (1 M in THF, 15.5 mL, 15.5 mmol) was added slowly and then warmed to ambient temperature, and the reaction mixture was stirred at ambient temperature for 2 h. The reaction was quenched with MeOM (5 mL), volatiles were evaporated and the residue was partitioned between water and EtOAc. The organic layer was washed with saturated NaHCO₃/water and brine, and then was dried (sodium sulfate) and evaporated to give compound 1 (1.26 g, 66.7%), MS (Cl) m/z 144.20 (MH⁺-Boc).

Step B

A solution of 1 (638 mg, 2.62 mmol) in THF (10 mL) was treated with 5-bromo-1-(2,6-difluorobenzyl)-6-methyluracil (869 mg, 2.62 mmol) and triphenylphosphine (1.03 g, 3.93 mmol) at ambient temperature, then di-tert-butylazodicarboxylate (906 mg, 3.93 mmol) was introduced. The reaction mixture was stirred at ambient temperature for 16 h and volatiles were evaporated. The residue was partitioned between saturated NaHCO₃/H₂O and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by flash chromatography (silica, 25% EtOAc/hexanes) to give compound 2 (1.39 g, 95.4%), MS (CI) m/z 456.10, 458.10 (MH⁺-Boc).

Step C

Compound 2 (1.0 g, 1.79 mmol) in benzene/EtOH/ethylene glycol dimethyl ether (20/2/22 mL) was added 2-fluoro-3-methoxyphenylboronic acid (382 mg, 2.24 mmol) and saturated Ba(OH)₂/water (0.5 M, 15 mL). The reaction mixture was deoxygenated with N₂ for 10 min, tetrakis(triphenylphosine)palladium (0) (208 mg, 0.18 mmol) was added and the reaction mixture was heated at 80° C. overnight under the protection of N₂. The reaction mixture was partitioned between brine and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by flash chromatography (silica, 30% EtOAc/hexanes) to give compound 3 (348 mg, 32.3%), MS (CI) m/z 502.20 (MH⁺-Boc).

Step D

A solution of 3 (300 mg, 0.5 mmol) in dichloromethane (2 mL) was added TFA (2 mL) and the reaction mixture was stirred at ambient temperature for 1 h. Volatiles were evaporated and the residue was partitioned between saturated NaHCO₃/water and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by reverse phase HPLC (C-18 column, 15-75% ACN/water) to give compound 4, MS (Cl) m/z 502.20 (MH³⁰ ).

By the above procedure, the compounds of the following Table 9 were also prepared.

TABLE 9

Cpd. NR₁R₂— MW No. R₅ R₆ (CR_(3a)CR_(3b))_(n)— —Q—R₄ (calc.) (obs.) 9-1  Me

485.5 486 9-2  Me

589.6 590 9-3  Me

526.6 527 9-4  Me

485.5 486 9-5  Me

513.5 514 9-6  Me

529.5 530 9-7  Me

512.5 513 9-8  Me

518.6 519 9-9  Me

532.6 533 9-10  Me

427 428 9-11  Me

505.6 506 9-12  Me

519.6 520 9-13  Me

520.6 521 9-14  Me

534.6 535 9-15  Me

532.6 533 9-16  Me

560.6 561 9-17  Me

534.6 535 9-18  Me

534.6 535 9-19  Me

529.6 530 9-20  Me

549.6 550 9-21  Me

554.6 555 9-22  Me

554.6 555 9-23  Me

575.6 576 9-24  Me

538.6 539 9-25  Me

537.6 538 9-26  Me

441.5 442 9-27  Me

546.6 547 9-28  Me

538.6 539 9-29  Me

579.6 447 9-30  Me

567.6 447 9-31  Me

533.6 534 9-32  Me

689.6 690 9-33  Me

590.7 591 9-34  Me

517.6 518 9-35  Me

666.7 667 9-36  Me

561.6 562 9-37  Me

574.6 575 9-38  Me

559.6 560 9-39  Me

643.7 644 9-40  Me

643.1 643 9-41  Me

561.7 562 9-42  Me

628.6 629 9-43  Me

666.7 667 9-44  Me

469.5 373 9-45  Me

505.5 373 9-46  Me

493.5 373 9-47  Me

459.5 373 9-48  Me

615.5 616 9-49  Me

516.6 517 9-50  Me

443.5 373 9-51  Me

592.6 593 9-52  Me

487.5 373 9-53  Me

500.5 501 9-54  Me

485.5 373 9-55  Me

569.6 372 9-56  Me

569.0 569 9-57  Me

487.6 373 9-58  Me

592.6 593 9-59  Me

495.5 399 9-60  Me

531.6 532 9-61  Me

519.5 399 9-62  Me

485.5 399 9-63  Me

641.5 642 9-64  Me

542.6 543 9-65  Me

469.5 470 9-66  Me

618.6 619 9-67  Me

513.5 514 9-68  Me

526.5 527 9-69  Me

511.6 512 9-70  Me

595.0 595 9-71  Me

513.6 399 9-72  Me

618.6 619 9-73  Me

493.6 397 9-74  Me

529.6 397 9-75  Me

517.6 397 9-76  Me

483.6 397 9-77  Me

639.6 640 9-78  Me

540.7 541 9-79  Me

467.6 468 9-80  Me

616.7 617 9-81  Me

511.6 512 9-82  Me

524.6 525 9-83  Me

509.6 510 9-84  Me

593.7 594 9-85  Me

593.1 593 9-86  Me

511.7 512 9-87  Me

578.6 579 9-88  Me

616.7 617 9-89  Me

509.5 413 9-90  Me

545.6 413 9-91  Me

533.6 413 9-92  Me

499.6 500 9-93  Me

556.6 557 9-94  Me

483.5 484 9-95  Me

632.7 633 9-96  Me

527.6 528 9-97  Me

540.6 541 9-98  Me

525.6 526 9-99  Me

527.6 528 9-100 Me

632.7 633 9-101 Me

554.5 555 9-102 Me

455.45 456 9-103 Me

581.66 9-104 Me

545.58 546 9-105 Me

588.65 9-106 Me

568.66 9-107 Me

572.62 9-108 Me

543.65 544.3 9-109 Me

506.55 507.2 9-110 Me

520.57 521.2 9-111 Me

552.61 553.3 9-112 Me

554.63 555.3 9-113 Me

570.63 571.3 9-114 Me

581.66 582.2 9-115 Me

525.98 526.2 9-116 Me

540.00 540.2 9-117 Me

525.98 526.2 9-118 Me

543.97 544.2 9-119 Me

560.42 561.1 9-120 Me

540.00 540.2 9-121 Me

574.45 574.0 9-122 Me

563.64 564.2 9-123 Me

497.58 498.2 9-124 Me

483.55 484.2 9-125 Me

469.52 470 9-126 Me

519.58 520.2 9-127 Me

520.57 521.5 9-128 Me

583.63 584.2 9-129 Me

535.58 536.2 9-130 Me

583.63 584.2 9-131 Me

501.57 502.2 9-132 Me

529.62 528 9-133 Me

556.60 557 9-134 Me

578.61 578 9-135 Me

540.60 541 9-136 Me

559.65 560.4 9-137 Me

547.64 548.5 9-138 Me

545.50 546.4 9-139 Me

585.62 586.4 9-140 Me

657.75 658.4 9-141 Me

561.66 562.6 9-142 Me

598.60 599.3 9-143 Me

598.60 599.3 9-144 Me

549.57 550.4 9-145 Me

639.59 640.4 9-146 Me

608.68 609.4 9-147 Me

607.65 608.2 9-148 Me

549.61 550.4 9-149 Me

485.54 486.4 9-150 Me

473.53 474 9-151 Me

471.39 472.2 9-152 Me

511.51 512.4 9-153 Me

583.65 584.2 9-154 Me

487.56 488.2 9-155 Me

524.49 525.4 9-156 Me

524.49 525.4 9-157 Me

527.62 528.4 9-158 Me

475.46 476.3 9-159 Me

565.48 566.4 9-160 Me

534.57 535.4 9-161 Me

533.55 534.5 9-162 Me

475.50 476.3 9-163 Me

499.55 500.4 9-164 Me

497.41 498.3 9-165 Me

537.53 538.4 9-166 Me

609.67 610.3 9-167 Me

513.58 514.6 9-168 Me

550.51 551.3 9-169 Me

550.51 551.2 9-170 Me

533.64 554.3 9-171 Me

501.48 502.3 9-172 Me

591.50 592.4 9-173 Me

560.59 561.3 9-174 Me

559.57 560.4 9-175 Me

501.52 502.3 9-176 Me

509.63 510.6 9-177 Me

497.62 498.5 9-178 Me

495.48 496.5 9-179 Me

535.60 536.6 9-180 Me

607.74 608.4 9-181 Me

511.65 512.5 9-182 Me

548.58 549.4 9-183 Me

551.71 552.4 9-184 Me

499.55 500.4 9-185 Me

589.57 590.5 9-186 Me

558.66 559.3 9-187 Me

557.64 558.3 9-188 Me

499.59 500.4 9-189 Me

525.59 526.4 9-190 Me

513.58 514.2 9-191 Me

511.44 512.5 9-192 Me

551.56 552.3 9-193 Me

623.69 624.4 9-194 Me

564.54 565.4 9-195 Me

564.54 565.4 9-196 Me

567.67 568.5 9-197 Me

515.51 516.3 9-198 Me

605.53 606.4 9-199 Me

574.6 575.4 9-200 Me

573.6 574.3 9-201 Me

515.6 516.3 9-202 Me

543.56 544.2 9-203 Me

609.07 609.2 9-204 Me

593.54 595.2 9-205 Me

498.62 499.3 9-206 Me

484.59 485.2 9-207 Me

595.64 596.4 9-208 Me

532.58 533.2 9-209 Me

532.58 533.2 9-210 Me

574.62 575 9-211 Me

Br 564.42 466/464 9-212 Me

Br 564.42 464/466 9-213 Me

575.69 576.3 9-214 Me

597.65 535.3 9-215 Me

597.65 598.2 9-216 Me

627.68 628.3 9-217 Me

517.57 518.2 9-218 Me

585.62 586.2 9-219 Me

617.69 618.2 9-220 Me

545.62 546.2 9-221 Me

576.68 577.3 9-222 Me

533.61 534.2 9-223 Me

491.53 492.2 9-224 Me

519.58 520.2 9-225 Me

622.71 623.3 9-226 Me

501.59 502.3 9-227 Me

460.49 461.2 9-228 Me

523.55 524.2 9-229 Me

553.57 554.2 9-230 Me

443.46 444.2 9-231 Me

511.51 512.2 9-232 Me

543.58 544.2 9-233 Me

429.44 430.1 9-234 Me

471.52 472.2 9-235 Me

502.57 503.3 9-236 Me

459.50 460.2 9-237 Me

417.42 418.1 9-238 Me

445.48 446.1 9-239 Me

548.60 549.2 9-240 Me

500.56 501.2 9-241 Me

527.60 528.3 9-242 Me

486.51 487.2 9-243 Me

549.57 550.2 9-244 Me

579.59 580.2 9-245 Me

469.48 470.2 9-246 Me

537.53 538.2 9-247 Me

569.60 570.2 9-248 Me

497.53 498.2 9-249 Me

528.59 529.2 9-250 Me

485.52 486.2 9-251 Me

443.44 444.1 9-252 Me

471.50 472.2 9-253 Me

574.62 575.2 9-254 Me

526.58 527.2 9-255 Me

525.68 526.3 9-256 Me

484.54 485.2 9-257 Me

547.64 548.3 9-258 Me

577.66 578.3 9-259 Me

467.55 468.2 9-260 Me

535.60 536.2 9-261 Me

567.67 568.3 9-262 Me

495.61 496.2 9-263 Me

526.66 527.3 9-264 Me

483.59 484.25 9-265 Me

441.51 442.2 9-266 Me

469.57 470.3 9-267 Me

572.69 573.3 9-268 Me

524.66 525.3 9-269 Me

541.63 542.3 9-270 Me

500.54 501.2 9-271 Me

563.59 564.2 9-272 Me

593.62 594.2 9-273 Me

483.51 484.2 9-274 Me

551.56 552.2 9-275 Me

583.63 584.2 9-276 Me

511.56 512.2 9-277 Me

542.62 543.3 9-278 Me

499.55 500.3 9-279 Me

457.47 458.2 9-280 Me

485.52 486.2 9-281 Me

588.65 589.3 9-282 Me

560.42 560 9-283 Me

539.66 540 9-284 Me

509.59 510 9-285 Me

510.60 511.5 9-286 Me

538.56 539.5 9-287 Me

516.58 517.4 9-288 Me

547.64 547 9-289 Me

519.56 534 9-290 Me

523.55 524.2 9-291 Me

615.65 616.3 9-292 Me

507.55 508.2 9-293 Me

522.56 523.6 9-294 Me

508.54 509.5 9-295 Me

537.57 538.7 9-296 Me

552.59 553.2 9-297 Me

538.56 539.5 9-298 Me

469.58 470.3 9-299 Me

484.59 485.3 9-300 Me

470.57 471.3 9-301 Me

546.65 547 9-302 Me

513.53 514 9-303 Me

495.56 496 9-304 Me

523.55 524 9-305 Me

537.57 538 9-306 Me

572.62 573 9-307 Me

537.57 538.3 9-308 Me

Br 505.36 505/507 9-309 Me

522.56 523 9-310 Me

505.56 506 9-311 Me

469.52 470 9-312 Me

505.56 506 9-313 Me

469.52 470 9-314 Me

519.58 520 9-315 Me

483.55 484 9-316 Me

519.58 520 9-317 Me

483.55 484 9-318 Me

534.60 535.3 9-319 Me

534.60 535.3 9-320 Me

511.56 512.5 9-321 Me

578.63 598 9-322 Me

427.44 428.1 9-323 Me

517.57 518.2 9-324 Me

518.56 519.2 9-325 Me

648.70 649.5 9-326 Me

561.66 562.5 9-327 Me

602.07 447.3 9-328 Me

491.53 447.4 9-329 Me

503.54 447.3 9-330 Me

519.58 447.2 9-331 Me

676.68 677.5 9-332 Me

604.65 605.3 9-333 Me

595.68 596.4 9-334 Me

632.70 633.4 9-335 Me

698.81 699.5 9-336 Me

574.62 575.4 9-337 Me

636.73 637.5 9-338 Me

574.59 575.4 9-339 Me

558.60 559.3 9-340 Me

487.56 488.3 9-341 Me

527.97 373.3 9-342 Me

417.42 373.1 9-343 Me

429.44 373.3 9-344 Me

445.48 373.2 9-345 Me

602.57 603.5 9-346 Me

530.54 531.3 9-347 Me

521.58 373.1 9-348 Me

558.60 559.3 9-349 Me

624.70 625.3 9-350 Me

442.43 373.3 9-351 Me

500.51 501.4 9-352 Me

562.63 563.4 9-353 Me

600.61 601.3 9-354 Me

584.62 585.2 9-355 Me

616.06 201.3 9-356 Me

513.58 399.2 9-357 Me

553.99 399.2 9-358 Me

443.44 399.3 9-359 Me

455.45 399.2 9-360 Me

471.50 399.3 9-361 Me

628.59 629.6 9-362 Me

556.56 557.3 9-363 Me

547.59 548.5 9-364 Me

584.62 585.2 9-365 Me

650.72 651.2 9-366 Me

468.45 399.1 9-367 Me

526.53 527.3 9-368 Me

588.65 589.5 9-369 Me

598.68 599.4 9-370 Me

582.69 583.4 9-371 Me

511.65 512.5 9-372 Me

552.06 397 9-373 Me

441.51 397.1 9-374 Me

453.53 397 9-375 Me

469.57 397.1 9-376 Me

626.66 627.6 9-377 Me

554.63 555.5 9-378 Me

545.67 546.4 9-379 Me

582.69 583.3 9-380 Me

648.79 649.6 9-381 Me

524.60 525.5 9-382 Me

586.72 587.5 9-383 Me

614.64 615.5 9-384 Me

598.64 599.4 9-385 Me

527.60 528.2 9-386 Me

568.01 568.5 9-387 Me

457.47 458 9-388 Me

485.52 486.3 9-389 Me

642.62 643.7 9-390 Me

570.59 571 9-391 Me

561.62 562.5 9-392 Me

598.64 599.4 9-393 Me

664.74 665.5 9-394 Me

540.56 541.6 9-395 Me

602.67 603.6 9-396 Me

442.43 373.3 9-397 Me

520.57 521.3 9-398 Me

520.57 521.2 9-399 Me

503.56 504.2 9-400 Me

532.58 533.2 9-401 Me

506.55 507 9-402 Me

506.55 507 9-403 Me

515.55 416 9-404 Me

531.6 532 9-405 Me

549.5 550 9-406 Me

550.57 550 9-407 Me

534.60 535 9-408 Me

534.60 535 9-409 Me

538.56 539 9-410 Me

524.54 525 9-411 Me

554.63 555 9-412 Me

H 335.35 336 9-413 Me

Br 533.41 533/535 9-414 Me

459.46 460 9-415 Me

H 454.51 455 9-416 Me

534.60 535.5 9-417 Me

520.57 521.5 9-418 Me

557.99 558 9-419 Me

539.55 540 9-420 Me

553.57 554 9-421 Me

537.57 538 9-422 Me

539.55 540 9-423 Me

553.57 554 9-424 Me

541.54 542 9-425 Me

541.54 542 9-426 Me

568.66 569 9-427 Me

664.14 664.2 9-428 Me

614.13 614.2 9-429 Me

590.04 590.2 9-430 Me

630.08 630.2 9-431 Me

469.48 470.2 9-432 Me

482.48 483.1 9-433 Me

466.52 467.2 9-434 Me

516.54 517.2 9-435 Me

595.68 596.3 9-436 Me

595.68 596.3 9-437 Me

538.56 539.2 9-438 Me

552.59 553.3 9-439 Me

506.55 507.2 9-440 Me

506.55 507.2 9-441 Me

520.57 521.2 9-442 Me

520.57 521.2 9-443 Me

537.57 538 9-444 Me

521.56 522.2 9-445 Me

521.56 522.2 9-446 Me

523.55 524.2 9-447 Me

523.55 524.2 9-448 Me

523.55 524.2 9-449 Me

530.57 531.2 9-450 Me

530.57 531.2 9-451 Me

530.57 531.2 9-452 Me

533.61 534.3 9-453 Me

533.61 534.3 9-454 Me

533.61 534.2 9-455 Me

535.58 536.2 9-456 Me

547.64 548.3 9-457 Me

548.63 549.3 9-458 Me

549.57 550.2 9-459 Me

535.58 536.2 9-460 Me

547.59 548.3 9-461 Me

556.00 556.2 9-462 Me

556.00 556.2 9-463 Me

556.00 556.2 9-464 Me

557.99 558.2 9-465 Me

557.99 558.2 9-466 Me

573.55 574.2 9-467 Me

544.59 545.2 9-468 Me

558.62 559.2 9-469 Me

495.52 496.2 9-470 Me

538.56 539 9-471 Me

495.52 496.2 9-472 Me

519.58 520.2 9-473 Me

519.58 520.2 9-474 Me

519.58 520.2 9-475 Me

521.56 535.2 9-476 Me

533.61 534.2 9-477 Me

535.58 536.2 9-478 Me

549.61 550.2 9-479 Me

551.65 552.2 9-480 Me

555.62 556.3 9-481 Me

552.59 553 9-482 Me

537.57 538.2 9-483 Me

539.55 540.2 9-484 Me

539.55 540.2 9-485 Me

541.54 542.2 9-486 Me

541.54 542.2 9-487 Me

541.54 542.2 9-488 Me

548.56 549.2 9-489 Me

548.56 549.3 9-490 Me

551.60 552.3 9-491 Me

551.60 552.2 9-492 Me

553.57 554.2 9-493 Me

553.57 554.2 9-494 Me

553.57 554.2 9-495 Me

565.58 566.2 9-496 Me

565.63 566.3 9-497 Me

565.63 566.3 9-498 Me

566.62 566.2 9-499 Me

567.56 567.3 9-500 Me

567.60 568.2 9-501 Me

569.64 568.2 9-502 Me

573.61 570.2 9-503 Me

573.99 574.2 9-504 Me

573.99 574.2 9-505 Me

573.99 574.2 9-506 Me

575.98 574.2 9-507 Me

575.98 576.2 9-508 Me

591.54 592.2 9-509 Me

562.58 563.2 9-510 Me

513.51 514.2 9-511 Me

513.51 514.2 9-512 Me

524.54 525.2 9-513 Me

547.64 548.3 9-514 Me

557.99 558.2 9-515 Me

525.63 526.3 9-516 Me

511.60 512.3 9-517 Me

523.55 524 9-518 Me

521.53 522 9-519 Me

555.63 556 9-520 Me

H 499.55 400 (MH- BOC)⁺ 9-521 Me

H 399.43 400 9-522 Me

H 397.42 398 9-523 Me

Br 478.33 478/480 9-524 Me

Br 476.31 476/478 9-525 Me

505.56 506.3 9-526 Me

519.58 520.3 9-527 Me

505.56 506.2 9-528 Me

519.58 520.2 9-529 Me

471.54 472.2 9-530 Me

485.57 486.3 9-531 Me

499.59 500.3 9-532 Me

521.60 522.2 9-533 Me

527.65 528.3 9-534 Me

539.66 540.3 9-535 Me

583.75 584.4 9-536 Me

523.62 524.3 9-537 Me

555.70 556.3 9-538 Me

483.55 484.2 9-539 Me

483.55 484.2 9-540 Me

497.58 498.3 9-541 Me

485.57 486.3 9-542 Me

499.59 500.3 9-543 Me

510.58 511.2 9-544 Me

513.62 514.3 9-545 Me

525.63 526.3 9-546 Me

501.54 502.2 9-547 Me

559.58 560.2 9-548 Me

515.57 516.2 9-549 Me

519.56 520.2 9-550 Me

557.99 558.2 9-551 Me

548.56 549.2 9-552 Me

541.54 542.2 9-553 Me

513.51 514.2 9-554 Me

543.60 544.2 9-555 Me

543.60 544.2 9-556 Me

529.58 530.1 9-557 Me

489.53 490.2 9-558 Me

557.65 558.2 9-559 Me

503.56 504.2 9-560 Me

545.64 546.2 9-561 Me

521.60 522.2 9-562 Me

537.57 538.2 9-563 Me

517.58 518.2 9-564 Me

559.66 560.2 9-565 Me

548.56 549.2 9-566 Me

515.57 516.2 9-567 Me

501.54 502.2 9-568 Me

515.57 516.2 9-569 Me

513.51 514.2 9-570 Me

529.58 530.2 9-571 Me

539.55 540.2 9-572 Me

557.65 558.3 9-573 Me

545.64 546.3 9-574 Me

503.56 504.3 9-575 Me

546.65 547.3 9-576 Me

559.66 560.3 9-577 Me

565.53 566.3 9-578 Me

548.56 549.2 9-579 Me

607.71 608.4 9-580 Me

505.53 506.2 9-581 Me

524.54 525.2 9-582 Me

538.56 539.2 9-583 Me

523.55 524.2 9-584 Me

523.55 524.2 9-585 Me

519.58 520.2 9-586 Me

535.58 536.2 9-587 Me

523.55 524.2 9-588 Me

521.56 522.2 9-589 Me

529.6 530.2 9-590 Me

531.61 532.3 9-591 Me

541.56 542.3 9-592 Me

513.51 514.2 9-593 Me

527.54 528.2 9-594 Me

601.74 602.4 9-595 Me

541.56 542.2 9-596 Me

543.62 542.2 9-597 Me

483.55 484.2 9-598 Me

471.54 472.1 9-599 Me

485.57 486.3 9-600 Me

499.59 500.3 9-601 Me

601.74 602.4 9-602 Me

527.54 528.2 9-603 Me

513.51 514.2 9-604 Me

546.63 547 9-605 Me

524.99 525 9-606 Me

501.54 502.2 9-607 Me

557.99 558.2 9-608 Me

541.54 542.2 9-609 Me

539.55 540.3 9-610 Me

601.74 602.4 9-611 Me

573.69 574.3 9-612 Me

545.64 546.3 9-613 Me

503.56 504.2 9-614 Me

541.61 542.3 9-615 Me

475.50 476.2 9-616 Me

489.53 490.3 9-617 Me

505.53 506.30 9-618 Me

526.55 527.2 9-619 Me

539.55 540.2 9-620 Me

539.55 540.2 9-621 Me

539.58 530.2 9-622 Me

608.47 608.1 9-623 Me

524.54 525.2 9-624 Me

559.53 560.2 9-625 Me

513.51 514.2 9-626 Me

530.56 531.2 9-627 Me

530.56 531.2 9-628 Me

592.40 594.1 9-629 Me

519.58 520.2 9-630 Me

521.58 522.2 9-631 Me

507.55 508.3 9-632 Me

525.54 526.2 9-633 Me

541.99 542.2 9-634 Me

537.57 538.3 9-635 Me

581.58 582.2 9-636 Me

551.60 552.3 9-637 Me

523.55 524.2 9-638 Me

575.55 576.2 9-639 Me

521.58 522.2 9-640 Me

573.55 574.2 9-641 Me

592.54 592.2 9-642 Me

629.67 630 9-643 Me

607.66 608 9-644 Me

643.70 644 9-645 Me

649.73 650 9-646 Me

647.66 648 9-647 Me

664.12 664 9-648 Me

671.71 672 9-649 Me

543.53 544.2 9-650 Me

524.54 525.2 9-651 Me

505.53 506.2 9-652 Me

513.51 514.2 9-653 Me

537.57 538.3 9-654 Me

513.51 514.2 9-655 Me

475.50 476.2 9-656 Me

503.56 504.3 9-657 Me

487.51 488.3 9-658 Me

501.54 502.2 9-659 Me

524.54 525.2 9-660 Me

543.53 544.2 9-661 Me

489.53 490.3 9-662 Me

541.56 542.3 9-663 Me

557.99 558.2 9-664 Me

526.55 527.2 9-665 Me

541.56 542.3 9-666 Me

559.53 560.2 9-667 Me

524.54 525.2 9-668 Me

513.51 514.2 9-669 Me

517.58 518.2 9-670 Me

524.54 525.2 9-671 Me

501.54 502 9-672 Me

639.66 540 9-673 Me

679.73 680 9-674 Me

659.70 660 9-675 Me

543.62 544.3 9-676 Me

543.62 544.3 9-677 Me

564.02 564.2 9-678 Me

531.61 532.3 9-679 Me

529.6 530.2 9-680 Me

539.55 540.2 9-681 Me

517.58 518.3 9-682 Me

537.57 538.2 9-683 Me

545.64 544.3 9-684 Me

539.55 540.2 9-685 Me

487.51 488.2 9-686 Me

609.77 610.3 9-687 Me

569.64 570.2 9-688 Me

531.61 532.3 9-689 Me

601.74 602.4 9-690 Me

557.99 558.2 9-691 Me

549.59 550.2 9-692 Me

517.58 518.2 9-693 Me

503.56 504.3 9-694 Me

503.56 504.3 9-695 Me

503.51 504 9-696 Me

537.53 538.2 9-697 Me

551.60 552.3 9-698 Me

529.6 530.2 9-699 Me

543.62 544.3 9-700 Me

529.6 530.2 9-701 Me

543.62 544.3 9-702 Me

523.55 524.2 9-703 Me

549.54 450 9-704 Me

503.56 504.3 9-705 Me

608.47 610.1 9-706 Me

529.58 530.2 9-707 Me

517.58 518.2 9-708 Me

503.56 504.3 9-709 Me

535.56 536.2 9-710 Me

489.53 490.2 9-711 Me

489.53 490.2 9-712 Me

503.56 504.2 9-713 Me

503.56 504.2 9-714 Me

489.53 490.2 9-715 Me

489.53 490.2 9-716 Me

523.55 524.2 9-717 Me

517.54 518.2 9-718 Me

523.55 524 9-719 Me

517.58 518.3 9-720 Me

535.57 536.3 9-721 Me

531.61 532.3 9-722 Me

503.56 504.3 9-723 Me

517.54 518 9-724 Me

543.60 544 9-725 Me

530.56 531 9-726 Me

553.57 554 9-727 Me

523.55 524.2 9-728 Me

509.52 510.2 9-729 Me

515.57 516.3 9-730 Me

529.6 530.3 9-731 Me

519.56 520.2 9-732 Me

487.519 488 9-733 Me

503.562 504 9-734 Me

517.6 518.2 9-735 Me

485.6 486.2 9-736 Me

541.6 542 9-737 Me

509.5 510.2 9-738 Me

491.5 492.2 9-739 Me

543.6 544.3 9-740 Me

515.6 516.3 9-741 Me

513.5 514 9-742 Me

637.8 638 9-743 Me

637.7 638 9-744 Me

625.7 626 9-745 Me

553.6 554 9-746 Me

661.6 662 9-747 Me

505.5 506.2 9-748 Me

519.5 520.2 9-749 Me

517.5 518 9-750 Me

489.5 490.2 9-751 Me

541.6 542 9-752 Me

536.5 537 9-753 Me

529.5 530 9-754 Me

542.6 543 9-755 Me

471.5 472.2 9-756 Me

485.5 486.2 9-757 Me

559.6 460.2 9-758 Me

527.6 428.2 9-759 Me

483.6 484.2 9-760 Me

511.6 512.2 9-761 Me

49.6 500.2 9-762 Me

497.6 498.2 9-763 Me

525.6 526.2 9-764 Me

533.5 534.2 9-765 Me

455.5 456.2 9-766 Me

455.5 456.2 9-767 Me

459.5 460.1 9-768 Me

459.5 459 9-769 Me

489.5 489 9-770 Me

487.5 488 9-771 Me

Br 442.3 442 9-772 Me

H 363.4 364 9-773 Me

587.6 588 9-774 Me

491.5 491

EXAMPLE 10 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 6-Methyl-5-(2-fluorophenyl)-oxaz-2,4-dione

To a stirred solution of 2′-fluorophenylacetone 1 (7.6 g. 50 mmol) in ether (50 mL) was added dropwise chlorosulfonylisocyanate (CSI, 16.2 g. 115 mmol) at room temperature. The yellow solution was stirred overnight, poured into ice (100 g) and basified with sodium carbonate. The product was extracted with ethyl acetate (2×200 mL) and the extract was washed with water and brine, dried over magnesium sulfate and concentrated in vacuo to give a yellow residue (9.5 g, proton NMR, about 70% product). The crude product was crystallized from ether-hexanes to give compound 2 as a yellow solid (3.6 g, 33% yield); ¹H NMR (CDCl₃): 2.14 (s, 3H), 7.16 (t, J=9.0 Hz, 1H), 7.24 (m, 2H), 7.41 (m, 1H), 9.20 (brs. 1H).

Step B 6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]oxaz-2,4-dione

DEAD (348 mg, 1.2 mmol) was added into a solution of oxazine 2 (221 mg, 1.0 mmol), triphenylphosphine (314 mg, 1.2 mmol) and N-Boc-(R)-phenylglysinol (249 mg, 1.05 mmol) in dry THF (5 mL). The mixture was stirred at room temperature for 2 hours, concentrated, and purified by chromatography on silica gel with 1:3 ethyl acetate/hexanes to give the product 3 (380 mg, 87%) as a white solid; ¹H NMR (CDCl₃): 1.39 (s, 9H), 2.14 (s, 3H), 4.02 (m, 1H), 4.28 (m, 1H), 5.21 (brs, 1H), 5.30 (m, 1H), 7.38 (m, 9H); MS (341, MH⁺-BuOCO).

Step C 6-Methyl-5-(2-fluorophenyl)-3-[2(R)-amino-2-phenylethyl]oxaz-2,4-dione Trifluoroacetic Acid Salt

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]oxaz-2,4-dione 3 (30 mg) was treated with trifluoroacetic acid (1 mL) at room temperature for 30 minutes. Concentration in vacuo gave the title compound 4 as a colorless oil in quantitative yield; ¹H NMR (CDCl₃): 2.05 & 2.08 (s, 3H), 4.10 (m, 1H), 4.45 (m, 1H), 4.62 (m, 1H), 7.15 (m, 3H), 7.40 (m, 6H), 8.20 (brs, 3H); MS: 341 (MH⁺).

Step D 6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]-1-(2-methoxybenzyl)uracil

A mixture of 6-methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]oxaz-2,4-dione 3 (29 mg) and 2-methoxybenzylamine (0.15 mL) was heated in a sealed reacti-vial at 100° C. for 1 hour. Chromatography on silica gel with 1:2 ethyl acetate-hexanes gave compound 5 as a colorless oil; ¹H NMR (CDCl₃): 1.40 (s, 9H), 2.04 (s, 3H), 3.87 (s, 3H), 4.18 (m, 1H), 4.44 (m, 1H), 5.22 (m, 2H), 5.65 (brs, 1H), 5.78 (m, 1H), 6.85-7.42 (m, 13H); MS: 460 (MH⁺-BuOCO).

The following protected intermediates were made using the same procedure but substituting different amines for 2-methoxybenzylamine. Acetic acid may be used to catalyze the reaction.

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]-1-(2,6-difluorobenzyl)uracil

¹H NMR (CDCl₃): 1.39 (s, 9H), 2.18 (s, 3H), 4.10 (m, 1H), 4.38 (m, 1H), 4.90-5.80 (m, 4H), 6.92 (m, 2H), 7.10-7.42 (m, 10H); MS: 466 (MH⁺-BuOCO).

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]-1-(2-chlorobenzyl)uracil

¹H NMR (CDCl₃): 1.40 (s, 9H), 2.02 (s, 3H), 4.15 (m, 1H), 4.50 (m, 1H), 5.35 (m, 3H), 5.62 (m, 1H), 6.95 (m, 13H); MS: 464 (MH⁺-BuOCO).

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]-1-(2-methylbenzyl)uracil

¹H NMR (CDCl₃): 1.40 (s, 9H), 2.02 (s, 3H), 2.37 (s, 3H), 4.15 (m, 1H), 4.42 (m, 1H), 5.72 (m, 1H), 6.80-7.42 (m, 13H); MS: 444 (MH⁺-BuOCO).

Step E 6-Methyl-5-(2-fluorophenyl)-3-[2(R)-amino-2-phenylethyl]-1-(2-methoxybenzyl)uracil Trifluoroacetic Acid Salt

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-tert-butoxycarbonylamino-2-phenylethyl]-1-(2-methoxybenzyl)uracil 5 (20 mg) was treated with trifluoroacetic acid (1 mL) at room temperature for 30 minutes. Concentration in vacuo gave the product 6 as a colorless oil in quantitative yield; ¹H NMR (CDCl₃): 2.04 (s, 3H), 3.82 & 3.85 (s, 3H), 4.20 (m, 1H), 4.62 (m, 2H), 5.10 (m, 2H), 6.82-7.40 (m, 13H), 8.05 (brs, 3H); MS: 460 (MH⁺).

The following products were also prepared using the same procedure.

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-amino-2-phenylethyl]-1-(2-chlorobenzyl)uracil Trifluoroacetic Acid Salt

¹H NMR (CDCl₃): 2.01 (s, 3H), 4.20 (m, 1H), 4.70 (m, 2H), 5.25 (m, 2H), 6.90-7.45 (m, 13H), 8.20 (brs, 3H); MS: 464 (MH⁺).

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-amino-2-phenylethyl]-1-(2-methylbenzyl)uracil Trifluoroacetic Acid Salt

¹H NMR (CDCl₃):2.00 (s, 3H), 2.27 & 2.34 (s, 3H), 4.15 (m, 4H), 4.62 (m, 2H), 5.15 (m, 2H), 6.80-7.40 (m, 13H); MS: 444 (MH⁺).

6-Methyl-5-(2-fluorophenyl)-3-[2(R)-amino-2-phenylethyl]-1-(2,6-difluorobenzyl)uracil Trifluoroacetic Acid Salt

¹H NMR (CDCl₃): 2.14 (s, 3H), 4.18 (m, 1H), 4.62 (m, 2H), 5.20 (m, 2H), 5.62 (brs, 3H), 6.85-7.40 (m, 13H); MS: 466 (MH⁺).

By the above procedure, the compounds of the following Table 10 were also prepared.

TABLE 10

MW Cpd. No. R₆ —Q—R₄ (calc.) (obs.) 10-1 

465.5 466 10-2 

393.5 394.2 10-3 

379.4 363 10-4 

407.5 323.3 10-5 

421.5 405.4 10-6 

435.5 436.2 10-7 

450.6 451.3 10-8 

433.5 417.3 10-9 

423.5 407.2 10-10

435.5 419.2 10-11

451.5 452.3 10-12

436.5 323.3 10-13

466.6 450.3 10-14

430.5 414.4 10-15

444.5 428.4 10-16

430.5 414.4 10-17

430.5 414.4 10-18

512.6 513.3 10-19

410.5 323.3 10-20

381.4 382.2 10-21

429.5 413.2 10-22

447.5 431.4 10-23

447.5 431.3 10-24

498.4 481.4 10-25

459.5 443.3 10-26

443.5 427.2 10-27

463.9 447.1 10-28

443.5 427.2 10-29

397.4 398.2 10-30

395.5 379.2 10-31

409.5 393.3 10-32

396.5 380.3 10-33

410.5 394.1 10-34

397.4 381.2 10-35

383.4 367.1 10-36

443.5 427.2 10-37

379.4 363.3 10-38

409.5 393.3 10-39

382.4 366.2 10-40

381.4 365.2 10-41

424.5 408.5 10-42

397.4 381.2 10-43

396.5 380.3 10-44

409.5 393.3 10-45

465.5 449.4 10-46

497.5 481.4 10-47

395.5 379.3 10-48

450.6 451.3 10-49

411.5 395.2 10-50

393.5 377.3 10-51

444.5 428.4 10-52

463.9 447.1 10-53

457.5 441.3 10-54

481.9 465.4 10-55

498.4 481.2 10-56

413.4 397.1 10-57

498.4 481.2 10-58

408.5 409.2 10-59

425.5 409.2 10-60

444.5 428.4 10-61

422.5 323.4 10-62

487.5 471.3 10-63

473.5 474.2 10-64

498.4 498.1 10-65

447.5 448.2 10-66

459.5 460.2 10-67

443.5 434.2 10-68

443.5 444.2 10-69

451.6 452.3 10-70

409.5 410.2 10-71

409.5 410.2 10-72

427.5 428.2

EXAMPLE 11 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-(1-ethoxyvinyl)-6-methyluracil

A solution of 1-(2,6-difluorobenzyl-3-[(2R)-tert-butylcarbonylamino-2-phenyl]ethyl-5-bromo-6-methyluracil 1 (500 mg, 0.91 mmol), tributyl(ethoxyvinyl)tin (0.39 mL) and (Ph₃P)₄Pd(0) (105 mg) in dioxane (5 mL) was heated at 100° C. under nitrogen for 2 hours. The reaction mixture was concentrated in vacuo and the crude product 2 was used for next step. MS: 442 (MH+-Boc).

Step B 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutyloxycarbonylamino-2-phenyl]ethyl-5-acetyl-6-methyluracil

A solution of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-(1-ethoxyvinyl)-6-methyluracil 2 (490 mg) in THF (10 mL) was treated with 2.5M aqueous HCl (3 mL) and stirred at r.t. for one hour. The reaction mixture was neutralized with NaHCO₃ and concentrated in vacuo to remove THF. The product was extracted with ethyl acetate. The extract was washed with water and brine, dried over MgSO₄ and concentrated in vacuo to give a brown solid. Chromatography on silica gel with 1:2 to 1:1 ethyl acetate/hexanes gave compound 3 as a white solid (227 mg, 50% yield); 1H NMR: 1.37 (s, 9H), 2.38 (s, 3H), 2.58 (s, 3H), 4.12 (dd, J=4.2, 10.0 Hz, 1H), 4.65 (dd, J=6.5, 10.0 Hz, 1H), 5.20 (m, 1H), 5.40 (d, J=12.0 Hz, 1H), 5.49 (d, J=12.0 Hz, 1H), 5.58 (d, J=6.0 Hz, 1H), 6.92 (t, J=8.0 Hz, 2H), 7.38 (m, 6H); MS: 414 (MH+-Boc).

Step C 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutoxycarbonylamino-2-phenyl]ethyl-5-(3-dimethylamino-1-oxopropenyl)-6-methyluracil

1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-acetyl-6-methyluracil 3 (44 mg) was suspended in DMFDMA (1.0 mL) and heated at 50° C. for 1 hour. The product was purified on silica gel with 1:1 ethyl acetate/hexanes to give compound 4 as a yellow oil; 1H NMR: 1.39 (s, 9H), 2.36 (s, 3H), 2.84 (s, 6H), 4.05 (m, 1H), 4.30 (m, 1H), 4.66 (d, J=12.0 Hz, 1H), 5.03 (m, 1H), 5.20 (d, J=12 Hz, 1H), 5.46 (d, J=12 Hz, 1H), 5.84 (d, J=7 Hz, 1H), 6.64 (d, J=12.0 Hz, 1H), 6.87 (t, J=8.0 Hz, 2H), 7.20-7.40 (m, 6H); MS: 596 (MH+).

Step D 1-(26-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(isoxazol-5-yl)-6-methyluracil

A mixture of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutoxycarbonylamino-2-phenyl]ethyl-5-(3-dimethylamino-1-oxopropenyl)-6-methyluracil 4 (95 mg), hydroxylamine hydrochloride (150 mg), sodium carbonate (18 mg) in methanol (5 mL) was acidified with acetic acid to pH˜4. The mixture was then heated at 120° C. for 1.5 hours, cooled down to r.t., filtered, and concentrated in vacuo to give the protected product. MS: 539 (MH+). The protected product was dissolved in dichloromethane (2 mL), treated with TFA (1 mL), and stirred at r.t. for 1 hour. Concentration in vacuo followed by purification on silica gel eluting with 1% aq. NH₄OH in ethyl acetate gave product 5; MS: 439 (MH+); 1H NMR (CD₃OD): 3.05 (s, 3H), 4.70 (m, 1H), 4.55 (m, 2H), 5.48 (d, J=12.0 Hz, 1H), 5.60 (d, J=12.0 Hz, 1H), 7.00 (t, J=8.0 Hz, 2H), 7.30-7.65 (m, 7H), 8.50 (d, J=6.0 Hz, 1H).

EXAMPLE 12 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-5 bromoacetyl-6-methyluracil

A solution of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-(1-ethoxyvinyl)-6-methyluracil 1 (3.68 g, 6.8 mmol) in THF (120 mL) and water (120 mL) was treated with N-bromosuccinimide (2.3 g) at r.t. and the mixture was stirred for 4 hours. THF was removed in vacuo and the product which precipitated on standing was collected by filtration and was washed with ether to give white solid 2 (1.6 g, 40%);1H NMR: 1.39 (s, 9H), 2.40 (s, 3H), 4.04 (dd, J=2.0, 7.0 Hz, 1H), 4.36 (d, J=7.0 Hz, 1H), 4.10 (d, J=5.5 Hz, 1H), 4.56 (d, J=5.5 Hz, 1H), 55.50 (m, 1H), 5.24 (d, J=12.0 Hz, 1H), 5.40 (brs, 1H), 5.50 (d, J=12.0 Hz, 1H), 6.94 (t, J=8.0 Hz, 1H), 7.36 (m, 6H); MS: 492 (MH+).

Step B 2-(2,6-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(5-methylthiazol-4-yl)-6-methyluracil

A solution of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-bromoacetyl-6-methyluracil (100 mg, 0.17 mmol) and thioacetamide (30 mg, 0.4 mmol) in ethanol (2 mL) was heated at 80° C. in a sealed reaction vessel for 3 hours. The reaction mixture was then concentrated in vacuo to give an oil and LCMS indicated protected product; MS: 569 (MH+). The protected product was dissolved in dichloromethane (2 mL) and treated with TFA (1 mL) at r.t. for 1 hour, and concentrated in vacuo. The product was purified on silica gel eluting with 5% aq. NH₄OH in ethyl acetate to give yellow solid 3; 1H NMR: 2.12 (s, 3H), 2.71 (s, 3H), 4.15-4.70 (m, 3H), 5.66 (s, 2H), 7.00 (t, J=8.0 Hz, 2H), 7.30 (m, 7H); MS: 469 (MH+).

Step C 1-(2,6-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(5-benzylaminolthiazol-4-yl)-6-methyluracil

A solution of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-bromoacetyl-6-methyluracil 2 (35 mg) and ammonium thioisocyanate (10 mg) in ethanol (1 mL) was heated at 80° C. in a sealed reaction vessel for 1 hour. Benzylamine (0.2 mL) was added and the mixture was heated at 80° C. overnight. The reaction mixture was then concentrated in vacuo, and the protected product was dissolved in dichloromethane (1 mL) and treated with TFA (1 mL) at r.t. for 1 hour. The mixture was concentrated in vacuo and the residue was purified on silica gel with 5% aq. NH₄OH in ethyl acetate to give product 4 as a yellow solid; ¹H NMR: 2.25 (s, 3H), 4.05 (dd, J=3.0, 7.5 Hz, 1H), 4.28 (dd, J=6.5, 7.5 Hz, 1H), 4.42 (m, 1H), 4.44 (s, 2H), 5.32 (d, J=12.0 Hz, 1H), 5.36 (d, J=12.0 Hz, 1H), 6.54 (s, 1H), 6.92 (t, J=8.0 Hz, 2H), 7.20-7.50 (m, 11H); MS: 560 (MH⁺).

Step D 1-(2,6-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(imidazolo[1,2-a]pyrid-2-yl)-6-methyluracil

A mixture of 1-(2,6-difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-bromoacetyl-6-methyluracil 2 (35 mg) and 2-aminopyridine (7 mg) in ethanol was heated at 80° C. overnight. The reaction mixture was then concentrated in vacuo, and the protected product was dissolved in dichloromethane (1 mL) and treated with TFA (1 mL) at r.t. for 1 hour. After concentration in vacuo, the product 5 was purified on preparative HPLC; 1H NMR: 2.32 (s, 3H), 4.04 (m, 1H), 4.67 (m, 2H), 5.17 (d, J=16.2 Hz, 1H), 5.41 (d, J=16.2 Hz, 1H), 6.92 (t, J=8.1 Hz, 2H), 7.24-7.40 (m, 7H), 7.73 (m, 1H), 7.80 (m, 1H), 8.03 (s, 1H), 8.30 (brs, 3H), 8.44 (d, J=5.5 Hz, 1H); MS: 488 (MH+).

TABLE 12

Cpd. MW No. —Q—R₄ (calc.) (obs.) 12-1 

468.5 469.1 12-2 

469.5 470.1 12-3 

497.6 498.2 12-4 

530.6 531.1 12-5 

544.6 545.2 12-6 

526.6 527.2 12-7 

488.5 489.2 12-8 

507.6 508.2 12-9 

508.6 509.1 12-10

575.6 576.2 12-11

545.6 546.2 12-12

563.6 564.2 12-13

590.6 591.1 12-14

559.6 560.2 12-15

487.5 488.2 12-16

539.6 540.2 12-17

559.6 560.2 12-18

573.7 574.2 12-19

509.5 510 12-20

598.6 599.2 12-21

565.0 565.2 12-22

565.0 565.1 12-23

583.0 583.1 12-24

548.6 549.2 12-25

559.6 560.2 12-26

575.6 576.2 12-27

605.7 606.3 12-28

573.7 574.2 12-29

573.7 574.2 12-30

573.7 574.2 12-31

573.7 574.2 12-32

559.6 560.2

EXAMPLE 13 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A. 5-Bromo-1-(2,6-difluorobenzyl)uracil

A suspension of 5-bromouracil (18.45 g, 96.6 mmol) in 300 mL of dichloroethane was treated with N,O-bis(trimethylsilyl)acetamide (48 mL, 39.5 g, 194 mmol). The reaction mixture was heated at 80° C. for 3 hr under the nitrogen. The solution was cooled down to ambient temperature, 2,6-difluorobenzyl bromide (25 g, 120 mmol) was added and the reaction mixture was heated at 80° C. overnight under the protection of nitrogen. The reaction was cooled down, quenched with MeOH (15 mL), and partitioned between dichloromethane (500 mL) and water (250 mL). The organic layer was washed with brine, dried (sodium sulfate), and evaporated to give a solid. The crude product was triturated with ether, filtered, and washed with ether three times to give compound 1 (15.2 g, 50%) as a white solid; MS (CI) m/z 316.90, 318.90 (MH⁺).

Step B 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-bromouracil

A solution of(R)-N-(tert-butoxycarbonyl)-2-phenylglycinol (14.97 g, 63.1 mmol) in anhydrous THF (300 mL) was treated with 5-bromo-1-(2,6-difluorobenzyl)uracil 1 (20 g, 63.1 mmol) and triphenylphosphine (20.68 g, 78.8 mmol) at ambient temperature, then diisopropylazodicarboxylate (15.52 mL, 15.94 g, 78.8 mmol) in THF (30 mL) was introduced via a dropping funnel. The reaction mixture was stirred at ambient temperature for 16 h and volatiles were evaporated. The residue was purified by flash chromatography (silica, 25% EtOAc/hexanes) to give compound 2 (31.15 g, 92.1%) as a white solid, MS (CI) m/z 436.0, 438.0 (MH⁺-Boc).

Step C 1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-5-(2,4,6-trimethylphenyl)uracil

To compound 2 (134 mg, 0.25 mmol) in toluene/H₂O/EtOH (6/3.75/0.75 mL) was added 2,4,6-trimethylphenyl boronic acid ester (87 mg, 1.5 eq), K₂CO₃ (86 mg, 2.5 eq), and saturated Ba(OH)₂/water (0.1 mL). The reaction mixture was deoxygenated with N₂ for 10 min, tetrakis(triphenylphosphine) palladium (0) (29 mg, 0.1 eq) was added and the reaction mixture was heated at 100° C. overnight under the protection of N₂. The reaction mixture was partitioned between brine and EtOAc. The organic layer was dried (sodium sulfate), evaporated, purified by flash chromatography (silica, 25% EtOAc/hexanes) to give compound 3 (130 mg) as a pale yellow oil.

Step D 1-(2,6-Difluorobenzyl-3-[(2R)-amino-2-phenyl]ethyl-5-(2,4,6-trimethylphenyl)uracil

TFA (3 mL) was added to a solution of 3 (130 mg, 0.22 mmol) in dichloromethane (3 mL) and the reaction mixture was stirred at ambient temperature for 2 hours. Volatiles were evaporated and the residue was partitioned between saturated NaHCO₃/water and EtOAc. The organic layer was dried (sodium sulfate), evaporated, and purified by prep TLC eluting with 5% MeOH in CH₂Cl₂ to give compound 4, MS (CI) m/z 476.2 (MH⁺).

TABLE 13

NR₁R₂— MW Cpd. No. (CR_(3a)CR_(3b))_(n)— —Q—R₄ (calc.) (obs.) 13-1

475.5 476.2 13-2

481.5 482 13-3

528.6 529 13-4

475.5 476.2

EXAMPLE 14 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 1-(2,6-Difluorobenzyl)-5-carbethoxyuracil

5-Carbethoxyuracil (5 g, 27.15 mmol) and N,O-bis(trimethylsilyl)acetamide (13.4 mL, 2 eq) in dichloroethane(35 mL) were heated at 80° C. for 2 hours. Difluorobenzyl bromide (8.4 g, 1.5 eq) was added and the reaction mixture was heated at 80° C. for 16 hours. The reaction was quenched with methanol and partitioned between methylene chloride and sodium bicarbonate solution. The organic layer was washed with brine, dried and concentrated in vacuo and the residue was triturated with ether to give compound 1 as a white solid (3.26 g).

Step B 1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-5-carbethoxyuracil

A solution of (R)-N-(tert-butoxycarbonyl)-2-phenylglycinol (316 mg, 1.33 mmol) in anhydrous THF (30 mL) was treated with 1-(2,6-difluorobenzyl)-5-carbethoxyuracil 1 (413 mg, 1.33 mmol) and triphenylphosphine (525 mg, 2 mmol) at ambient temperature, then diisopropylazodicarboxylate (460 mg, 2 mmol) in THF (5 mL) was introduced via a dropping funnel. The reaction mixture was stirred at ambient temperature for 5 h and volatiles were evaporated. The residue was purified by flash chromatography (silica, 35% EtOAc/hexanes) to give compound 2 (427 mg) as a white foam.

Step C 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-n-butylamidouracil

A solution of triethylaluminum (1.9 M in toluene, 0.26 mL, 0.5 mmol) was added to n-butylamine (0.1 mL, 1 mmol) in dichloroethane and the reaction mixture was sealed under nitrogen and stirred for ½ hour. 1-(2,6-Difluorobenzyl-3-[(2R)-tertbutylcarbonylamino-2-phenyl]ethyl-5-carbethoxyuracil 2 was added and the mixture was stirred at 70-80° C. for 12 hours to give 3. Trifluoroacetic acid (1 mL) was added and the reaction mixture was stirred for 1 hour. The mixture was concentrated in vacuo and the residue was partitioned between methylene chloride and sodium carbonate solution. The organic layer was washed with brine, dried and concentrated to give a residue which was purified by prep HPLC to give compound 4 (56 mg, MH⁺457).

TABLE 14

Cpd. NR₁R₂— MW No. (CR_(3a)CR_(3b))_(n)— —Q—R₄ (calc.) (obs.) 14-1

456.5 457.2 14-2

456.5 457.2 14-3

456.5 457.2 14-4

413.4 414.1 14-5

523.6 424.2 14-6

423.5 424.2

EXAMPLE 15 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-5-bromo-6-ethyluracil

1-(2,6-Difluorobenzyl-3-[(2R)-tert-butoxycarbonylamino-2-phenyl]ethyl-5-bromo-6-methyluracil 1 (550 mg, 1 mmol) was dissolved in THF (10 mL) and the solution was cooled to 0° C. Lithium bis(trimethylsilyl)amide (1.0 M in THF, 1.3 mL, 1.3 mmol) was added dropwise and the reaction was stirred for 40 minutes at 0° C. Iodomethane (0.093 mL, 1.5 mmol) was added dropwise and after 30 minutes, water was added and the mixture extracted with ethyl acetate. Concentration in vacuo gave compound 2 as a yellow foam.

TABLE 15

Cpd. NR₁R₂— MW No. (CR_(3a)CR_(3b))_(n)— —Q—R₄ (calc.) (obs.) 15-1

509.5 510.2 15-2

491.5 492 15-3

534.6 535 15-4

H 428.5 429 15-5

504.6 505 15-6

546.65 15-7

548.58 549.2 15-8

503.6 504.3 15-9

523.6 524

EXAMPLE 16 SYNTHESIS OF REPRESENTATIVE COMPOUNDS

Step A 1-(2,6-Difluorobenzyl)-3-(4-methyl-2R-guanidopentyl)-5-(2-fluoro-3-methoxyphenyl)-6-methyluracil

A solution of 1-(2,6-difluorobenzyl)-3-(4-methyl-2R-aminopentyl)-5-(2-fluoro-3-methoxyphenyl)-6-methyluracil 1 (75 mg), (1H)-pyrazole-1-carboxamidine hydrochloride (23 mg) diisopropylethylamine (21 mg) in anhydrous DMF was heated at 40-50° C. overnight (0.5 mL). The reaction mixture was treated with water and the product was extracted with ethyl acetate. The extract was dried over MgSO₄, filtered and concentrated in vacuo and the residue was purified on silica gel (Et₃N/MeOH/CHCl₃ (2:5:93) as elutant) to give white solid 2. MS: 518 (MH⁺).

It will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without departing from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims. 

We claim:
 1. A compound having the following structure:

or a stereoisomer or pharmaceutically acceptable salt thereof, wherein: Q is a direct bond or —(CR_(8a)R_(8b))_(r)—Z—(CR_(10a)R_(10b))_(s)—; A is S or NR₇; r and s are the same or different and independently 0, 1, 2, 3, 4, 5 or 6; n is 2, 3 or 4; Z is a direct bond or —O—, —S—, —NR₉—, —SO—, —SO₂—, —OSO₂—, —SO₂O—, —SO₂NR₉—, —NR₉SO₂—, —CO—, —COO—, —OCO—, —CONR₉—, —NR₉CO—, —NR₉CONR_(9a), —OCONR₉— or —NR₉COO—; R₁ and R₂ are the same or different and independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl, substituted heterocyclealkyl, —C(R_(1a))(═NR_(1b)), or —C(NR_(1a)R_(1c))(═NR_(1b)); or R₁ and R₂ taken together with the nitrogen atom to which they are attached form a heterocyclic ring or a substituted heterocyclic ring; R_(3a) and R_(3b) are the same or different and, at each occurrence, independently hydrogen, alkyl, substituted alkyl, alkoxy, alkylthio, alkylamino, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl, substituted heterocyclealkyl, —COOR₁₄ or —CONR₁₄R₁₅; or R_(3a) and R_(3b) taken together with the carbon atom to which they are attached form a homocyclic ring, substituted homocyclic ring, heterocyclic ring or substituted heterocyclic ring; or R_(3a) and R_(3b) taken together form ═NR_(3c); or R_(3a) and the carbon to which it is attached taken together with R₁ and the nitrogen to which it is attached form a heterocyclic ring or substituted heterocyclic ring; R₄ is higher alkyl, substituted alkyl, aryl, substituted aryl, heterocycle, substituted heterocycle, —COR₁₁, —COOR₁₁, —CONR₁₂R₁₃, —OR₁₁, —OCOR₁₁, —OSO₂R₁₁, —SR₁₁, —SO₂R₁₁, —NR₁₂R₁₃, —NR₁₁COR₁₂, —NR₁₁CONR₁₂R₁₃, —NR₁₁SO₂R₁₂ or —NR₁₁SO₂NR₁₂R₁₃; R₅ is hydrogen, halogen, lower alkyl, substituted lower alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, alkoxy, alkylthio, alkylamino, cyano or nitro; R₆ is higher alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl; R₇ is hydrogen, —SO₂R₁₁, cyano, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl; and R_(1a), R_(1b), R_(1c), R_(3c), R_(8a), R_(8b), R₉, R_(9a), R_(10a), R_(10b), R₁₁, R₁₂, R₁₃, R₁₄ and R₁₅ are the same or different and, at each occurrence, independently hydrogen, acyl, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl or substituted heterocyclealkyl; or R_(1a) and R_(1b), R_(8a) and R_(8b), R_(10a) and R_(10b), R₁₂ and R₁₃, or R₁₄ and R₁₅ taken together with the atom or atoms to which they are attached form a homocyclic ring, substituted homocyclic ring, heterocyclic ring or substituted heterocyclic ring.
 2. The compound of claim 1 wherein A is S.
 3. The compound of claim 1 wherein A is NR₇.
 4. The compound of claim 1 wherein R₁ is aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocycle, substituted heterocycle, heterocyclealkyl or substituted heterocyclealkyl.
 5. The compound of claim 4 wherein heterocycle is heteroaryl, substituted heterocycle is substituted heteroaryl, heterocyclealkyl is heteroarylalkyl, and substituted heterocyclealkyl is substituted heteroarylalkyl.
 6. The compound of claim 5 wherein R₁ is heteroarylalkyl or substituted heteroarylalkyl.
 7. The compound of claim 1 wherein R₁ is phenylalkyl or substituted phenylalkyl.
 8. The compound of claim 1 wherein R₁ is benzyl.
 9. The compound of claim 1 wherein R₁ is hydrogen or lower alkyl.
 10. The compound of claim 1 wherein R₂ is hydrogen, alkyl or substituted alkyl.
 11. The compound of claim 1 wherein R₂ is hydrogen or methyl.
 12. The compound of claim 1 wherein Q is a direct bond.
 13. The compound of claim 1 wherein Q is —(CR_(8a)R_(8b))_(r)—Z—(CR_(10a)R_(10b))_(s)—.
 14. The compound of claim 1 wherein R_(3a) and R_(3b) are, at each occurrence, hydrogen.
 15. The compound of claim 1 wherein R_(3a) is hydrogen, alkyl, aryl or arylalkyl.
 16. The compound of claim 1 wherein R_(3a) is hydrogen, methyl, isobutyl, cyclohexyl, phenyl or benzyl.
 17. The compound of claim 1 wherein R_(3b) is, at each occurrence, hydrogen.
 18. The compound of claim 1 wherein n is
 1. 19. The compound of claim 1 wherein n is
 2. 20. The compound of claim 19 wherein —(R_(3a)R_(3b)C)_(n)— has the structure —C(R_(3a))(R_(3b))CH₂.
 21. The compound of claim 20 wherein R_(3a) is benzyl.
 22. The compound of claim 20 wherein R_(3a) is alkyl.
 23. The compound of claim 22 wherein R_(3a) is isobutyl or cyclohexyl.
 24. The compound of claim 20 wherein R_(3b) is hydrogen or methyl.
 25. The compound of claim 1 wherein R₄ is substituted aryl or substituted heterocycle.
 26. The compound of claim 1 wherein R₄ is substituted phenyl.
 27. The compound of claim 26 wherein R₄ is phenyl substituted with halogen, alkoxy, or both halogen and alkoxy.
 28. The compound of claim 1 wherein R₅ is H, lower alkyl or substituted lower alkyl.
 29. The compound of claim 1 wherein R₅ is hydrogen or methyl.
 30. The compound of claim 1 wherein R₆ is aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl.
 31. The compound of claim 1 wherein R₆ is arylalkyl, substituted arylalkyl, heteroarylalkyl or substituted heteroarylalkyl.
 32. The compounds of claim 1 wherein R₆ is benzyl or substituted benzyl.
 33. The compound of claim 1 wherein R₆ is benzyl substituted with two halogens.
 34. The compound of claim 1 wherein Q is a bond and R₄ is substituted aryl or heterocycle.
 35. The compound of claim 1 wherein R₁ is —CH₂(heteroaryl) or —CH₂CH₂(heteroaryl).
 36. The compound of claim 1 wherein R₁ and R₂ taken together with the nitrogen atom to which they are attached form a heterocyclic ring or substituted heterocyclic ring.
 37. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier or diluent. 